1.Validation of UHPLC-MS/MS-Network Pharmacology Joint Experiment to Explore the Mechanism of Action of Tongluo Muzu Powder in Improving Cartilage Injury
Di TIAN ; Zixiu LIU ; Likai YU ; Zishan SU ; Shangqi LIU ; Mingqing FENG ; Zeen WANG ; Nongshan ZHANG
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(3):363-374
OBJECTIVE To study and analyse the main components of Tongluo Muzu Powder transdermal absorption solution,as well as to investigate its mechanism of action in the treatment of osteoarthritis of the knee.METHODS The transdermal absorption solution of Tongluo Muzu Powder was extracted by Franz vertical diffusion cell,and its components were analyzed by UHPLC-MS/MS.Then TCMSP,Swiss target prediction,Gene Cards and other databases were introduced to predict the possible targets of the effec-tive components of transdermal absorption solution for the prevention and treatment of knee osteoarthritis,and Cytoscape software was used to construct the"drug-component-disease-target"visual network,and then the protein interaction network was obtained through the string database to find the core targets.Finally,AutoDock and PyMOL were used to verify the molecular docking of main active in-gredients and targets,and the gene ontology(GO)function analysis of core genes and the enrichment analysis of Kyoto Encyclopedia of genes and genomes(KEGG)pathway were conducted.Primary mouse chondrocytes were extracted,and the predicted targets of net-work pharmacology were verified by ELISA,Western blot,qPCR,etc.RESULTS A total of 48 effective components of the transder-mal absorption solution of Tongluo Muzu Powder and 88 possible targets for the treatment of knee osteoarthritis were screened out.The results of enrichment analysis showed that it was mainly involved in the process of apoptosis and oxidative stress.Molecular docking suggested that the main active ingredients and targets had good binding ability.The results showed that the lyophilized powder of the transdermal absorption solution of Tongluo Muzu Powder could reduce the concentration of inflammatory factors in the supernatant of chondrocytes after LPS intervention in a dose-dependent manner(P<0.05,P<0.01).The protein expression of p-PI3K/PI3K,p-AKT/AKT,MMP13 and p53 was decreased,and the protein expression of collagen Ⅱ was increased(P<0.05,P<0.01).At the same time,the mRNA expression of MMP13 and p53 was decreased and the mRNA expression of collagen Ⅱ was increased(P<0.05,P<0.01).In addition,it reduced the fluorescence intensity of TUNEL staining in cells(P<0.01).CONCLUSION By combining UH-PLC-MS/MS technology with network pharmacology,the main active ingredients of Tongluo Muzu Powder transdermal absorption solu-tion are preliminarily understood,and its potential mechanism of action in the treatment of knee osteoarthritis are predicted.
2.Validation of UHPLC-MS/MS-Network Pharmacology Joint Experiment to Explore the Mechanism of Action of Tongluo Muzu Powder in Improving Cartilage Injury
Di TIAN ; Zixiu LIU ; Likai YU ; Zishan SU ; Shangqi LIU ; Mingqing FENG ; Zeen WANG ; Nongshan ZHANG
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(3):363-374
OBJECTIVE To study and analyse the main components of Tongluo Muzu Powder transdermal absorption solution,as well as to investigate its mechanism of action in the treatment of osteoarthritis of the knee.METHODS The transdermal absorption solution of Tongluo Muzu Powder was extracted by Franz vertical diffusion cell,and its components were analyzed by UHPLC-MS/MS.Then TCMSP,Swiss target prediction,Gene Cards and other databases were introduced to predict the possible targets of the effec-tive components of transdermal absorption solution for the prevention and treatment of knee osteoarthritis,and Cytoscape software was used to construct the"drug-component-disease-target"visual network,and then the protein interaction network was obtained through the string database to find the core targets.Finally,AutoDock and PyMOL were used to verify the molecular docking of main active in-gredients and targets,and the gene ontology(GO)function analysis of core genes and the enrichment analysis of Kyoto Encyclopedia of genes and genomes(KEGG)pathway were conducted.Primary mouse chondrocytes were extracted,and the predicted targets of net-work pharmacology were verified by ELISA,Western blot,qPCR,etc.RESULTS A total of 48 effective components of the transder-mal absorption solution of Tongluo Muzu Powder and 88 possible targets for the treatment of knee osteoarthritis were screened out.The results of enrichment analysis showed that it was mainly involved in the process of apoptosis and oxidative stress.Molecular docking suggested that the main active ingredients and targets had good binding ability.The results showed that the lyophilized powder of the transdermal absorption solution of Tongluo Muzu Powder could reduce the concentration of inflammatory factors in the supernatant of chondrocytes after LPS intervention in a dose-dependent manner(P<0.05,P<0.01).The protein expression of p-PI3K/PI3K,p-AKT/AKT,MMP13 and p53 was decreased,and the protein expression of collagen Ⅱ was increased(P<0.05,P<0.01).At the same time,the mRNA expression of MMP13 and p53 was decreased and the mRNA expression of collagen Ⅱ was increased(P<0.05,P<0.01).In addition,it reduced the fluorescence intensity of TUNEL staining in cells(P<0.01).CONCLUSION By combining UH-PLC-MS/MS technology with network pharmacology,the main active ingredients of Tongluo Muzu Powder transdermal absorption solu-tion are preliminarily understood,and its potential mechanism of action in the treatment of knee osteoarthritis are predicted.
3.Clinical progress of inflatable video-assisted mediastinoscopic transhiatal esophagectomy
Shangqi SONG ; Yang HU ; Yuyang XU ; Zheng LIU ; Weipeng HU ; Longqi CHENG ; Yong YUAN
Chinese Journal of Digestive Surgery 2023;22(4):474-480
Surgery is an important method for the treatment of resectable esophageal cancer. With the continuous development of minimally invasive esophageal technology, video-assisted thoracoscopic esophagectomy (VATE) has demonstrated obvious advantages over conventional open surgery and has been widely accepted. However, there are still some esophageal cancer patients who cannot benefit from VATE. Inflatable video-assisted mediastinoscopic transhiatal esophagectomy(IVMTE) does not require one-lung ventilation, reduces postoperative complications, expands surgical indications, and brings surgical opportunities for patients with impaired lung function and thoracic lesions, which has become a new choice for the treatment of esophageal cancer. However, the limited field of surgical view and the tunneling surgical approach undoubtedly increase the difficulty of surgery, and how to clearly expose the anatomical structures and thoroughly dissect the lymph nodes has always been the key points and difficulties of surgery. The authors review relevant literatures to discuss the clinical progress and limitations of IVMTE.
4.The effect of perioperative use of antibiotics on clinical indicators of thoracic surgery: A case control study
LIAO Hu ; SONG Shangqi ; PU Qiang ; MEI Jiandong ; XIAO Zhilan ; XIA Liang ; LIU Lunxu
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2018;25(5):393-396
Objective To explore the effect of standardized use of antibiotics on clinical indicators after thoracic surgery, such as pulmonary infection rate, incision infection rate, average length of hospital stay and total hospitalization cost. Methods We selected 468 patients (an observation group) who were hospitalized and received thoracic surgery from August to October 2011, 3 months after the implementation of the preventive antibiotics use protocol for thoracic surgery in West China Hospital, Sichuan University, and selected 343 patients (a control group) in the same period of the previous year (from August to October 2010). There were 326 males and 142 females with a mean age of 52.0±15.5 years in the observation group, and 251 males and 92 females with a mean age of 51.4±15.9 years in the control group. The level of antibiotic use, medication time, antibiotics cost, postoperative incision infection, incidence of pulmonary infection, postoperative hospital stay and total hospitalization cost were compared between the two groups. Results Compared with the control group, the time for preventive use of antibiotics was significantly shorter in the observation group (3.6±2.4 d vs. 6.1±3.1 d, P=0.020) and the total cost of antibiotic use significantly reduced (1 230.0±2 151.0 yuan vs. 2 252.0±1 764.0 yuan, P<0.001). There was no significant difference between the two groups in hospitalization cost (36 345.0±13 320.0 yuan vs. 35 821.0±11 991.0 yuan, P=0.566), postoperative hospital stay (10.6±8.4 d vs. 10.7±5.3 d, P=0.390), the incidence of postoperative wound infection or postoperative pulmonary infection (1.5% vs. 2.3%, P=0.430; 19.2% vs. 22.2%, P=0.330). Conclusion The standardized use of antibiotics in thoracic surgery does not cause postoperative pulmonary infection and incision infection, and has no negative impact on clinical indicators. Significantly reducing the level of antibiotics use may have a positive effect on reducing medication time, in-hospital infection and the incidence of drug-resistant strains.
5.Effects of immunosuppressants on co-stimulated lymphocyte's Th1/Th2 cytokine production
Shangqi YANG ; Xiaoda TANG ; Xiao GU ; Yong LIU ; Peijun ZHOU ; Zhichen GUAN
Chinese Journal of Urology 2008;29(5):337-341
Objective To analyze the effect of cyclosporin A(CsA), rapamycin(RPM) and macophenolic acid(MPA) on the co-stimulated lymphocytes, CD28 and CD40, and their production of Th1/Th2 cytokine, IL-2, IFN-γ, IL-4, IL-10 and IL-12. Methods The experimental groups were divided into ①mono-stimulating and co-stimulating groups: CD3 mAb mono-stimulating (group a),CD3 mAb+CD28 mAb co-stimulating (group b), CD3 mAb+CD28 mAb+CD40 L mAb co-stimulating(group c), CD3 mAb+CD28 mAb+CTLA4 mAb co-stimulating (group d). ②CsA groups: 300 ng/ml of CsA was added to group a, b, c and d. ③RPM groups: 300 ng/ml of RPM was added to group a,b, c and d. ④MPA groups:300 ng/ml of MPA was added to group a, b, c and d. The cytokine production was measured by ELISA.Results The co-stimulated CD28 and CD40 Th1/Th2 cytokines production of IFN-γ, IL-2, IL-4 and IL-10 were significantly increased. Compared with group a, IFN-γ increased from (248.91±11.20)ng/ml to (555.08±24.42)ng/ml and (548.19±33.06)ng/ml, IL-2 increased from (29.48±8.61)ng/ml to (1100.82±99.29)ng/ml and (842.23±29.31)ng/ml, IL-4 increased from (32.29±6.76)ng/ml to (116.02±15.03)ng/ml and (147.28±18.07)ng/ml, IL-10 increased from (147.01±10.47)ng/ml to (291.79±12.47)ng/ml and (302.52±35.18)ng/ml,respectively, P<0. 01. Compared group b with group c, the Th1 cytokines production was decreased.IL-2 and IL-12 decreased (P<0.05). The Th2 cytokine IL-4 production was increased (P<0. 05).CTLA4 mAb and three other immunosuppressants, CsA, RPM and MPA, inhibited co-stimulated lymphocyte's both cytokines Th1 and Th2 production. The inhibitory effect of CsA on Th1/Th2 cytokine production was more significant than RPM and MPA did. The co-inhibitory effect of CTLA4 mAb and CsA was observed as well. The increased co-stimulated CD28 and CD40 IL-12 production could be suppressed by MPA. CsA and RPM had no inhibitory effect on the IL-12 production.Conclusions CD28/CD40 co-stimulatory pathway plays the key role in lymphocyte activation and Th1/Th2 cytokine production. CsA, RPM and MPA can inhibit co-stimulated lymphocyte's Th1 and Th2 cytokine production. CsA and CTLA4 mAb have co-inhibitory effect on co-stimulated lymphocyte's Th1/Th2 cytokines production. CD40 L mAb decreases the Th1 cytokines production(including IL-12) and increases the Th2 (mainly IL-4) production, which may be the mechanism of its longevity effect on allograft.

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