1.23-Hydroxybetulinic Acid Inhibits NNK and LPS-induced Lung Inflammation-related Carcinogenesis by Regulating Immune Cells in Mice
Penghaobang LIU ; Wenbin DUAN ; Yajuan CHEN ; Lanying CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):98-106
ObjectiveTo investigate the suppressive effect of 23-hydroxybetulinic acid (23-HBA), a key constituent of Pulsatillae Radix, on the pulmonary inflammation-related carcinogenesis induced by the combined exposure of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and lipopolysaccharide (LPS) in mice, alongside exploring its influence on immune cells and delving into the underlying mechanisms. MethodsA murine model of pulmonary inflammation-related carcinogenesis induced by NNK combined with LPS was established. Mice were randomly assigned into blank control, model, aspirin (10 mg·kg-1), and low-, medium-, and high-dose (3.75, 7.5, 15 mg·kg-1, respectively) 23-HBA groups. The treatment lasted for 26 weeks, after which the spleen, lung, and peripheral blood samples were collected. Lung and spleen indices were calculated. Histopathological changes in the lung tissue were observed by hematoxylin-eosin staining. Immunohistochemistry was employed to assess the expression levels of thyroid transcription factor-1 (TTF-1), neuron-specific enolase (NSE), and proliferating cell nuclear antigen (Ki-67) in the lung tissue. High-throughput protein microarray was employed to measure the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in the mouse serum. Flow cytometry was employed to evaluate the expression of macrophages, myeloid-derived suppressor cells (MDSCs), and exhausted T lymphocytes in the lung and spleen tissue. Molecular docking was performed to predict the binding affinity of 23-HBA to Janus kinase 2 (JAK2), Src homology 2 domain-containing phosphatase 2 (SHP2), and suppressor of cytokine signaling 3 (SOCS3). Western blot was performed to assess the protein levels of phosphorylated-signal transducer and activator of transcription 3 (p-STAT3), p53, and SHP2 in the M1-activated macrophages and A549 lung adenocarcinoma cells treated with 23-HBA. ResultsCompared with the normal group, the lung and spleen indexes of the model group were increased to varying degrees (P<0.05, P<0.01), the expression of TTF-1, NSE and Ki-67 protein was significantly increased (P<0.05, P<0.01), and the serum levels of TNF-α, IL-1β and IL-6 were significantly increased (P<0.01). The number of macrophages in the model group was significantly decreased (P<0.01), and the number of exhausted T cells and MDSCs was significantly increased (P<0.05, P<0.01). Compared with the model group, the spleen and thymus index of mice in each dose group of 23-HBA decreased significantly (P<0.05), and the lung index of mice in the middle dose group of 23-HBA decreased significantly (P<0.05). The high and middle dose groups of 23-HBA could improve the occurrence of inflammatory infiltration and malignant lesions in the lungs of mice induced by NNK combined with LPS in the model group. The expression of TTF-1 in the middle and high dose groups of 23-HBA was significantly lower than that in the model group (P<0.05, P<0.01). The expression of NSE and Ki-67 protein in each dose group of 23-HBA was significantly lower than that in the model group (P<0.05, P<0.01). The contents of IL-1β in the low and high dose groups of 23-HBA were significantly decreased (P<0.05), and the contents of IL-6 and TNF-α in each dose of 23-HBA were significantly decreased (P<0.05, P<0.01). The number of macrophages in the lung of the middle dose group of 23-HBA was significantly increased (P<0.05), and the number of exhausted T cells and MDSCs expressing PD-1 in the lung was significantly decreased (P<0.05, P<0.01). In addition, 23-HBA had strong molecular docking ability to SHP2, SOCS3 and JAK2 (≥7 kcal·mol-1), and significantly down-regulated the protein levels of p-STAT3, SHP2 and p53 in M1 macrophages and A549 lung adenocarcinoma (P<0.01). Conclusion23-HBA holds promise as a potential therapeutic agent for mitigating pulmonary inflammation and inhibiting malignant transformation induced by the combination of LPS and NNK. It may exert effects by regulating immune cell responses, improving the tumor immune microenvironment, and regulating key signaling pathways.
2.Effect of Huayu Tongluo moxibustion on learning-memory ability in rats with vascular dementia based on hippocampal Mst1/NF-κB p65 pathway.
Ping WANG ; Jun YANG ; Yu KONG ; Yating ZHANG ; Yinqiu FAN ; Haiping SHI ; Lanying LIU
Chinese Acupuncture & Moxibustion 2025;45(1):53-60
OBJECTIVE:
To observe the effects of Huayu Tongluo (transforming stasis and unblocking collaterals) moxibustion on learning-memory ability and hippocampal mammalian sterile 20-like kinase 1 (Mst1)/nuclear factor κB (NF-κB) p65 pathway related to inflammatory response in rats with vascular dementia (VD).
METHODS:
A total of 60 male Wistar rats of SPF grade were randomly divided into a sham operation group (12 rats) and a modeling group (48 rats). VD model was established by the method of modified bilateral common carotid artery permanent ligation in the modeling group. Thirty-six rats with successful modeling were randomly divided into a model group, a moxibustion group and a western medication group, with 12 rats in each group. Huayu Tongluo moxibustion was applied at "Dazhui" (GV14), "Baihui" (GV20) and "Shenting" (GV24) in the moxibustion group, 20 min each time, once a day, 7 day-intervention was as one course, and 1 day-interval was taken between two courses, for a total of 3 courses. In the western medication group, piracetam was given 0.72 mg/kg by intragastric administration, twice a day, the course of intervention was same as that of the moxibustion group. The learning-memory ability was detected by Morris water maze test; the morphology of hippocampal CA1 region was observed by HE staining; the mRNA expression of Mst1, M1 microglia markers CD86, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) was detected by real-time PCR; the levels of IL-6 and TNF-α in hippocampus were detected by ELISA; and the protein expression of Mst1 and NF-κB p65 in hippocampus was detected by Western blot in rats of each group.
RESULTS:
Compared with the sham operation group, the escape latency was prolonged in the model group (P<0.05); compared with the model group, the escape latency was shortened in the moxibustion group and the western medication group (P<0.05). The cells in the CA1 region of hippocampus were disordered, cell collapse and irregular nuclei could be observed in the model group; compared with the model group, the cell arrangement in the CA1 region of hippocampus was more regular, and the damage was improved in the moxibustion group and the western medication group. Compared with the sham operation group, the mRNA expression of Mst1, CD86, IL-6 and TNF-α, as well as the protein expression of Mst1, NF-κB p65 in hippocampus were increased in the model group (P<0.05). Compared with the model group, the mRNA expression of Mst1, CD86, IL-6 and TNF-α, as well as the protein expression of Mst1, NF-κB p65 in hippocampus were decreased in the moxibustion group and the western medication group (P<0.05). Compared with the sham operation group, the levels of IL-6 and TNF-α in hippocampus were increased in the model group (P<0.05). Compared with the model group, the levels of IL-6 and TNF-α in hippocampus were decreased in the moxibustion group and the western medication group (P<0.05).
CONCLUSION
Huayu Tongluo moxibustion can improve the learning-memory ability of VD rats, the mechanism may be related to regulating the activation of microglia through Mst1/NF-κB p65 pathway, reducing the release of pro-inflammatory factors i.e. IL-6 and TNF-α, so as to alleviating the damage of inflammatory factors in the hippocampus of VD rats.
Animals
;
Male
;
Rats
;
Moxibustion
;
Hippocampus/metabolism*
;
Rats, Wistar
;
Dementia, Vascular/genetics*
;
Memory/drug effects*
;
Humans
;
Transcription Factor RelA/genetics*
;
Learning
;
Protein Serine-Threonine Kinases/genetics*
;
Acupuncture Points
;
Interleukin-6/genetics*
;
Signal Transduction/drug effects*
;
Drugs, Chinese Herbal
3.Correlation between the skin surface temperature at the related back-shu points and the pulmonary ventilation function in patients with chronic persistent asthma based on the theory of "lung governing the skin and hair".
Shaoqian ZHAO ; Mengyu FU ; Nanxin HUANG ; Jipeng ZHOU ; Jinglin HUANG ; Wei LIU ; Hesheng WANG ; Lanying LIU
Chinese Acupuncture & Moxibustion 2025;45(3):274-279
OBJECTIVE:
To observe the skin surface temperature at the related back-shu points in the patients with the different levels of pulmonary ventilation function in chronic persistent asthma, and to explore the correlation between the skin temperature at the back-shu points and pulmonary ventilation function indexes based on "lung governing the skin and hair".
METHODS:
Sixty-one patients with chronic persistent asthma, based on the level of pulmonary ventilation function, were assigned into a reduced pulmonary ventilation function group (reduced function group, 32 cases) and a normal pulmonary ventilation function group (normal function group, 29 cases). In the two groups, the skin surface temperature was measured in the sites of bilateral Feishu (BL13), Geshu (BL17), Pishu (BL20) and Shenshu (BL23); and the pulmonary ventilation function indexes (the percentage of predicted value of forced vital capacity [FVC%pred], the percentage of predicted value of forced expiratory volume in the first second [FEV1%pred], the percentage of predicted value of FEV1/FVC [FEV1/FVC%pred] and the percentage of predicted value of the peak expiratory flow [PEF%pred]) were recorded. The correlation between the skin surface temperature of acupoints and pulmonary ventilation function was analyzed.
RESULTS:
Compared with the normal function group, the surface skin temperature at the bilateral Feishu (BL13), Geshu (BL17), Pishu (BL20) and Shenshu (BL23) was higher in the reduced function group (P<0.05, P<0.01). Compared with the normal function group, FEV1%pred, FEV1/FVC%pred and PEF%pred were decreased in the reduced function group (P<0.001). There was no significant difference in FVC%pred between the two groups (P>0.05). The skin surface temperature at the bilateral Feishu (BL13), Geshu (BL17), Pishu (BL20) and Shenshu (BL23) was negatively correlated with FVC%pred, FEV1%pred, FEV1/FVC%pred and PEF%pred in 61 patients with chronic persistent asthma (P<0.001, P<0.01, P<0.05).
CONCLUSION
The skin surface temperature at back-shu points is elevated in line with the the decline of pulmonary ventilation function in the patients with chronic persistent asthma, presenting a negative correlation with pulmonary ventilation function indexes. It is preliminarily verified that back-shu point is characterized by reflecting the visceral disorders.
Humans
;
Female
;
Male
;
Asthma/therapy*
;
Middle Aged
;
Adult
;
Skin Temperature
;
Lung/physiopathology*
;
Acupuncture Points
;
Pulmonary Ventilation
;
Aged
;
Chronic Disease/therapy*
;
Young Adult
;
Hair
4.Thoughts on the Formulation of"2023 Guidelines for Diagnosis and Treatment of Depression with Integrative Chinese and Western Medicine"
Liyuan GUO ; Jianjun WANG ; Zhen WANG ; Lanying LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(1):12-18
The integration of Chinese and Western Medicine(ICWM)in the diagnosis and treatment of depression has been a long-standing issue in the field of mental health.In order to maximize the benefits of this combined approach,the"2023 Guidelines for Diagnosis and Treatment of Depression with Integrative Chinese and Western Medicine"(GDTD-ICWM)has been developed based on organizing evidence-based and standardized workflows for ICWM.This article aims to discuss the background,highlights,summary and outlook of the"Guidelines",as well as offer operational suggestions to guide clinical practices in order to strengthen the implementation,practice and promotion of the"Guidelines".
5.Comparative Study on the Topological Attributes of Deficiency and Excess Pattern of Cerebral Small Vessel Disease Mild Cognitive Impairment Based on fMRI Graph Theory Network
Jianjun WANG ; Weimin YANG ; Haotao ZHENG ; Jianxiang CHEN ; Dongbin CAI ; Hanqing LYU ; Xiude QIN ; Lanying LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(1):69-75
Objective This study aimed to explore the differences in global and local brain network topological properties between deficient pattern(DP)and excess pattern(EP)of mild vascular cognitive impairment caused by subcortical small vessel disease based on graph theory network.Methods Patients were recruited prospectively and were classified with DP and EP subtype.The global small-world topological attributes and local nodes were calculated for the comparison of DP,EP,and healthy controls(CN)using the GRETNA platform.Results The three groups all had small-world attributes,but only the patients in EP had a significantly lower small world attribute δ in the range of 0.05-0.26 than the control group(P<0.05).The node efficiency and node strength indicators of multiple brain region were able to significantly distinguish the DP group from the EP group.However,there was no positive brain region in the node efficiency of the DP patients(P>0.05),and only a few brain regions showed increased node strength efficiency(P<0.05).Conclusion The results indicate that the syndrome of DP and EP have significantly different neuroimaging phenotypes,providing a basis for further research of biological classification based on Chinese Medicine syndromes.
6.Efficacy and Safety of Acupuncture in Treating Major Depressive Disorders:A Systematic Review and Meta Analysis
Wangtao LI ; Mingqi TU ; Liyuan GUO ; Yiyin HU ; Haomin SUN ; Peiqi LI ; Lanying LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(1):76-87
Objective To evaluate the effectiveness and safety of acupuncture in patients with Major Depressive Disorders.Methods Search the database for relevant literature published before April 15,2023.Select randomized controlled trial that meeting the enrollment criteria,evaluate the quality of the literature and classify before meta-analysis systematic evaluation.Results 15 RCTs are included in this study,with a total of 1077 patients.The results showed that acupuncture treatment,compared with antidepressants alone,The score of Hamilton Depression Scale(HAMD)in patients with liver-qi depression disorder[MD=-0.73,95%CI(-1.22,-0.24),P<0.05],the score of TCM Syndrome Scale[MD=-1.89,95%CI(-2.35,-1.44),P<0.001],effective rate[RR=1.09,95%CI(1.03,1.153),P=0.001]and TESS score[MD=-2.76,95%CI(-3.68,-1.85),P<0.001]were statistically significant.Compared with fluoxetine hydrochloride,the effective rate of HAMD scale score was statistically significant[RR=1.07,95%CI(1.01,1.14),P<0.05].Conclusion Compared with antidepressants alone,acupuncture therapy has the advantages for the improvement of HAMD score in patients,the improvement of Traditional Chinese medicine symptoms,and significantly less adverse effects than those in the control group.Through this study,it is considered that acupuncture can be a safe and effective treatment modality.However,due to the quantity and quality of the included literature,the above conclusions still need to be verified by more high-quality clinical trials.
7.Identification of Dalbergia odorifera and Its Counterfeits by HS-GC-MS
Li ZHAO ; Xiaowei MENG ; Jiarong LI ; Qing ZHU ; Xianwen WEI ; Ronghua LIU ; Lanying CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(2):156-163
ObjectiveTo screen the differential markers by analyzing volatile components in Dalbergia odorifera and its counterfeits, in order to provide reference for authentication of D. odorifera. MethodThe volatile components in D. odorifera and its counterfeits were detected by headspace gas chromatography-mass spectrometry(HS-GC-MS), and the GC conditions were heated by procedure(the initial temperature of the column was 50 ℃, the retention time was 1 min, and then the temperature was raised to 300 ℃ at 10 ℃ for 10 min), the carrier gas was helium, and the flow rate was 1.0 mL·min-1, the split ratio was 10∶1, and the injection volume was 1 mL. The MS conditions used electron bombardment ionization(EI) with the scanning range of m/z 35-550. The compound species were identified by database matching, the relative content of each component was calculated by the peak area normalization method, and principal component analysis(PCA), orthogonal partial least squares-discrimination analysis(OPLS-DA) and cluster analysis were performed on the detection results by SIMCA 14.1 software, and the differential components of D. odorifera and its counterfeits were screened out according to the variable importance in the projection(VIP) value>2 and P<0.05. ResultA total of 26, 17, 8, 22, 24 and 7 volatile components were identified from D. odorifera, D. bariensis, D. latifolia, D. benthamii, D. pinnata and D. cochinchinensis, respectively. Among them, there were 11 unique volatile components of D. odorifera, 6 unique volatile components of D. bariensis, 3 unique volatile components of D. latifolia, 6 unique volatile components of D. benthamii, 8 unique volatile components of D. pinnata, 4 unique volatile components of D. cochinchinensis. The PCA results showed that, except for D. latifolia and D. cochinchinensis, which could not be clearly distinguished, D. odorifera and other counterfeits could be distributed in a certain area, respectively. The OPLS-DA results showed that D. odorifera and its five counterfeits were clustered into one group each, indicating significant differences in volatile components between D. odorifera and its counterfeits. Finally, a total of 31 differential markers of volatile components between D. odoriferae and its counterfeits were screened. ConclusionHS-GC-MS combined with SIMCA 14.1 software can systematically elucidate the volatile differential components between D. odorifera and its counterfeits, which is suitable for rapid identification of them.
8.Construction of a therapeutic effect prediction model for bronchial asthma
Qidong HUANG ; Minxi LI ; Yilong LI ; Wanqi SHAO ; Shumei ZHAO ; Xiaoyan GONG ; Lindu ZHAO ; Lanying LIU
Journal of Beijing University of Traditional Chinese Medicine 2024;47(5):729-740
Objective We aimed to investigate the feasibility and method of constructing a traditional Chinese medicine(TCM)curative effect prediction model based on the data of Kechuanting acupoint plastering therapy in the treatment of bronchial asthma(asthma).Methods Data from the Chronic Disease Management Research Platform of 303 patients with asthma who were treated with Kechuanting acupoint plastering therapy for 6 weeks in the Department of Acupuncture and Rehabilitation of Jiangsu Hospital of Traditional Chinese Medicine from June to August 2018 to 2021 were selected.We used Phyton 3.10 for statistical analysis.After data preprocessing,the influencing factors were used to build models by Logistic regression,support vector machine,K-means clustering algorithm,Bayes algorithm,random forest method and Light gradient boosting machine(LightGBM)respectively,with the improvement of asthma control test score(ACT),forced expiratory volume in one second(FEV1)and exhaled nitric oxide(FeNO)as the outcome indicators.Then,the models were compared and analyzed.Subsequently,the superior model was used to establish the efficacy prediction model and verify its stability to obtain the accuracy rate and eliminate the relatively important factors.Results The accuracy rate of the Kechuanting acupoint plastering therapy curative effect prediction model established by the LightGBM model was more than 70%.Five important factors were selected,including allergic history,tabacco and alcohol abuse,plastering duration,ACT before treatment,and FeNO before treatment.According to the classification analysis and the relationship between the important factors and the outcome indicators,Kechuanting acupoint plastering therapy significantly improved the ACT of patients with no history of allergy,no tabacco and alcohol abuse,and poor ACT:5-15 points(P<0.05).Furthermore,Kechuanting acupoint plastering therapy improved FeNO more significantly in patients with more than 3 years of treatment than those with no more than 3 years(P<0.05).However,Kechuanting acupoint plastering therapy only improved FeNO in a few patients with poor asthma control levels(P<0.05)and severe airway inflammation(FeNO>50×10-9)(P>0.05).Conclusion Acupoint plastering application has a significant effect on improving the control level of asthma,but its effect on improving airway inflammation is limited.It is feasible to use data from the chronic disease management research platform to construct the prediction model.After optimization and testing,the predictive model established based on the data of this study may provide an effective evaluation tool for targeted clinical treatment.
9.Distribution and significance of CD1a+and CD83+dendritic cells in lung tissue of COPD mice
Lanying ZHANG ; Fuan ZHANG ; Maomao LIU ; Jie CHEN ; Jian ZHOU ; Yuting LIU ; Yao OUYANG
Tianjin Medical Journal 2024;52(9):913-916
Objective To study the distribution and significance of CD1a and CD83 positive dendritic cells(DCs)in lung tissue of chronic obstructive pulmonary disease(COPD)mice.Methods Twenty C57BL/6 mice were randomly divided into the air control group and the smoked COPD group(n=10 for each group).COPD mouse model was established using cigarette smoking method.Mice were executed within 24 h after the last cigarette smoking,and right lower lung was collected.Body mass changes and lung histopathological changes of mice were observed in two groups.Mean linear intercept(MLI)was measured,and expression levels of CD1a+and CD83+DCs in lung tissue were detected by immunohistochemistry.Results The body mass of mice at 7,14,21 and 28 d after modeling was lower in the smoked COPD group than that in the air control group(P<0.05).HE staining showed that the normal alveolar structure of lung tissue of mice in the smoked group was disrupted,with multiple alveoli fused with each other to form a larger alveolar lumen,a large number of inflammatory cells infiltrated in alveolar intervals,and walls of the alveoli were thickened.COPD modeling was successful.Compared with the air control group,MLI values(μm)increased in the smoked COPD group(28.30±3.47 vs.50.40±3.60),and the number of CD1a+DCs(per field of view)in lung tissue increased(9.58±2.18 vs.17.08±3.67),while the number of CD83+DCs(per field of view)decreased(19.78±4.95 vs.8.02±3.30)(all P<0.05).Conclusion The number of CD1a+DCs in lung tissue is increased and the number of CD83+DCs in lung tissue is decreased in the smoked COPD group of mice,and cigarette smoking may have impaired DC maturation.
10.Effect of Jogging on Efficacy in Patients with Depressive Disorder with Liver Depression and Spleen Deficiency
Yiying HU ; Wei LI ; Hui ZHANG ; Liyuan LUO ; Lanying LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(6):1440-1445
Objective To explore the effect of jogging on the patients with depression disorder of liver-stagnation and spleen-deficiency.Methods 100 patients with liver-stagnation and spleen-deficiency depression disorder were randomly selected from Tongde Hospital of Zhejiang Province from March to September 2022.They were divided into observation group and control group(50 cases each)by random number table method.The control group received conventional antidepressant drug treatment,and the observation group was given jogging exercise regimen on the basis of the control group:exercise at least 4 times a week,30-60 min each time,and the heart rate was maintained at 110-120 times/min for 4 weeks.The depressive disorder of the patients was assessed by Zung's Self-rating Depression Scale(SDS)and 17-item Hamilton Depression Scale(HAMD-17).Results After jogging exercise intervention,the total effective rate of the observation group was 94%,which was significantly higher than that of the control group 80%(P<0.05).HAMD-17 score was significantly lower in the observation group than in the control group after intervention(F=12.575,P<0.05).The scores of cognitive impairment,block and sleep disturbance in the observation group were significantly lower than those in the control group after intervention(F=12.575,P<0.05).The body weight factor scores of the observation group and the control group were significantly higher after intervention than before intervention(P>0.05).Conclusions Jogging exercise intervention therapy can significantly improve the depression of patients with liver-qi and spleen-deficiency depression disorder,and jogging can be an effective method to prevent and treat depression disorder.

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