1.Combining label-free quantitative proteomics and 2D-DIGE to identify the potential targets of Sini Decoction acting on myocardial infarction.
Fei FENG ; Weiyue ZHANG ; Yan CAO ; Diya LV ; Yifeng CHAI ; Dandan GUO ; Xiaofei CHEN
Chinese Journal of Natural Medicines (English Ed.) 2025;23(8):1016-1024
Sini Decoction (SNT) is a traditional formula recognized for its efficacy in warming the spleen and stomach and dispersing cold. However, elucidating the mechanism of action of SNT remains challenging due to its complex multiple components. This study utilized a synergistic approach combining two-dimensional fluorescence difference in gel electrophoresis (2D-DIGE)-based drug affinity responsive target stability (DARTS) with label-free quantitative proteomics techniques to identify the direct and indirect protein targets of SNT in myocardial infarction. The analysis identified 590 proteins, with 30 proteins showing significant upregulation and 51 proteins showing downregulation when comparing the SNT group with the model group. Through the integration of 2D-DIGE DARTS with proteomics data and pharmacological assessments, the findings indicate that protein disulfide-isomerase A3 (PDIA3) may serve as a potential protein target through which SNT provides protective effects on myocardial cells during myocardial infarction.
Myocardial Infarction/genetics*
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Proteomics/methods*
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Drugs, Chinese Herbal/chemistry*
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Animals
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Protein Disulfide-Isomerases/genetics*
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Male
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Two-Dimensional Difference Gel Electrophoresis/methods*
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Humans
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Rats
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Rats, Sprague-Dawley
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Electrophoresis, Gel, Two-Dimensional
2.Effects of Pingan qushi prescription on traditional Chinese medicine syndrome,adverse events,and compliance in young and middle-aged patients with hypertension
Songbo CHAI ; Yakang DU ; Shujuan ZHANG ; Jiangbo WU ; Xiaofei WANG
Journal of China Medical University 2024;53(1):8-14
Objective To investigate the effects of the Pingan qushi prescription on traditional Chinese medicine syndrome,adverse events,and compliance in young and middle-aged hypertensive patients.Methods One hundred thirty-two young and middle-aged patients with hypertension were selected and randomly divided into two groups.After elimination,64 cases were included in each group.For 2 months,the control group was treated with irbesartan tablets,and the observation group underwent the Pingan qushi prescription based on the control group.The blood pressure,curative effect,traditional Chinese medicine syndrome score,safety,adverse events,and prognosis-related indicators were compared between the two groups.Results The total effective rate of the observation group(96.88%,92.19%)was higher than that of the control group(82.52%,78.13%),and the difference was statistically significant(P = 0.005,P = 0.025).After 1 and 2 months of treatment,the blood pressure,syndrome scores,ALD,PRA,AngⅡ,and ET-1 in the observation group were lower than those in the control group,and the differences were statistically significant(all P<0.05).There was no sig-nificant difference in the incidence of adverse reactions between the two groups(P>0.05).There was no statistically significant difference(P>0.05)in the incidence of adverse events during treatment between the observation group(1.56%)and control group(10.94%).Conclusion The treatment of young and middle-aged community patients with hypertension undergoing the Pingan qushi pre-scription can further improve patients'symptoms,increase blood pressure control,promote prognosis improvement.
3.Efficacy of Fuyang Tongluo therapy combined with conventional western medicine in elderly patients with coronary heart disease and heart failure
Songbo CHAI ; Yakang DU ; Shujuan ZHANG ; Jiangbo WU ; Xiaofei WANG
Journal of China Medical University 2024;53(6):531-535
Objective To explore the therapeutic efficacy of the Fuyang Tongluo therapy combined with conventional western medicine in elderly patients with heart failure due to coronary heart disease.Methods In this single-blind randomized controlled trial,96 elderly patients with coronary heart disease and heart failure were randomly divided into three groups,each consisting of 32 individuals.Control group A was treated with conventional western medicine(atorvastatin combined with metoprolol),control group B with Fuyang Tongluo therapy,and the observation group with Fuyang Tongluo therapy and conventional western medicine.The three groups were compared in terms of therapeutic efficacy and traditional Chinese medicine symptom points as well as blood interleukin(IL)-23 and IL-17 levels,left ventricular end-systolic diameter(LVESD),left ventricular end-diastolic diameter(LVEDD),and left ventricular ejection fraction(LVEF)values before and after treatment.Results The total clinical efficacy rate in the observation group was 93.75%,which was significantly higher than those in control groups A(68.75%)and B(65.63%)(P<0.05).After 4 and 8 weeks of treatment,the primary and minor symptom scores,total symptom scores,LVESD,LVEDD,and IL-23 and IL-17 levels in the observation group were significantly lower than those in control groups A and B(P<0.05).Conclusion The combination of the Fuyang Tongluo therapy with conventional western medicine exhibited good therapeutic efficacy in elderly patients with coronary heart disease and heart failure.These effects were achieved by inhibiting excessive expression of the inflammatory axis factors IL-23 and IL-17 and promoting the recovery of cardiac function.
4.Clinical Observation of 32 Cases of Malignant Lymphoma Treated with Brentuximab Vedotin Combined with Chemotherapy
Xiaofei CHAI ; Honghan QIAO ; Lei ZHANG ; Xin LI ; Ling LI ; Xinhua WANG ; Xiaorui FU ; Xudong ZHANG ; Zhenchang SUN ; Feifei NAN ; Mingzhi ZHANG
Cancer Research on Prevention and Treatment 2023;50(3):288-292
Objective To retrospectively evaluate the clinical efficacy and safety of brentuximab vedotin(BV) combined with chemotherapy in the treatment of malignant lymphoma. Methods We collected the data of 32 lymphoma patients with CD30-positive status, including 14 cases of Hodgkin's lymphomas, 2 cases of diffuse large B-cell lymphomas, and 16 cases of mature T/NK cell lymphomas. Chemotherapy combined with BV was administered to all patients for a minimum of two cycles. The efficacy of the treatment was evaluated according to Lugano criteria every two cycles. Results Complete response rate and overall response rate after four cycles of treatment were 22% and 50%, respectively. Sixteen cases (50.0%) had grades 1 and 2 toxicity, and 16 cases (50.0%) had grade 3 toxicity or higher. The most common adverse events were neutropenia (50.0%), pneumonia (46.9%), and anemia (43.8%). The most common grade 3 or higher adverse events were pneumonia (18.8%) and febrile neutropenia (12.5%). Four patients discontinued brentuximab vedotin because of severe adverse events. Conclusion BV is effective in treating relapsed and refractory CD30- positive Hodgkin's lymphoma and peripheral T-cell lymphoma, and its overall safety is acceptable.
5.Clinical observation of ticagrelor combined with Compound xueshuantong capsule in the treatment of unstable angina pectoris
Songbo CHAI ; Yakang DU ; Shujuan ZHANG ; Jiangbo WU ; Weiping ZHANG ; Xiaofei WANG
China Pharmacy 2023;34(23):2901-2905
OBJECTIVE To observe the efficacy and safety of ticagrelor combined with Compound xueshuantong capsules in the treatment of unstable angina pectoris. METHODS Totally 120 patients with unstable angina pectoris with deficiency of Qi and Yin combined admitted to Sanmenxia Hospital of Traditional Chinese Medicine from January 2021 to December 2022 were randomly divided into clopidogrel group (group A), ticagrelor group (group B) and combined medication group (group C), with 40 patients in each group. In addition to conventional treatment, group A was given clopidogrel orally; group B was given ticagrelor orally; group C was given ticagrelor and Compound xueshuantong capsule orally. After 12 weeks of treatment, the clinical efficacy, frequency of angina attacks, coagulation function indicators, cardiac function indicators, traditional Chinese medicine syndrome efficacy, and the incidence of adverse cardiovascular events and bleeding incidence were evaluated in the 3 groups. RESULTS The total effective rates of group A, group B and group C were 77.5%, 85.0% and 90.0%, respectively. The incidence of adverse cardiovascular events and bleeding events were 7.5%, 7.5% and 5.0% in the respective groups, with no statistically significant difference (P>0.05). The frequency of angina attacks, duration of angina attacks and duration of dyspnea were significantly reduced or shortened in all 3 groups after treatment compared to before treatment (P<0.05). The frequency of angina attacks in group C was significantly lower than that in groups A and B after treatment (P<0.05). The levels of fibrinogen (FBG) and D-dimer in all 3 groups were significantly lower after treatment compared to before treatment (P<0.05); group A had significantly higher levels of FBG and D-dimer compared to group B and C (P<0.05). The left ventricular end-diastolic diameter and left ventricular end-systolic diameter in all three groups were significantly shorter after treatment, and the left ventricular ejection fraction was significantly higher compared to before treatment (P<0.05), but there were no statistically significant differences among those groups (P>0.05). The total effective rates of traditional Chinese medicine syndrome efficacy in groups A, B and C were 67.5%, 80.0% and 87.5%, respectively, with group C being significantly higher than group A (P<0.05). CONCLUSIONS In addition to conventional treatment, ticagrelor combined with Compound xueguantong capsules can more significantly reduce the frequency of angina attacks in patients with unstable angina pectoris, reduce the levels of FBG and D- dimer, improve traditional Chinese medicine syndrome efficacy, and do not increase the risk of bleeding.
6.Screening potential P-glycoprotein inhibitors by combination of a detergent-free membrane protein extraction with surface plasmon resonance biosensor.
Yuhong CAO ; Jiahao FANG ; Yiwei SHI ; Hui WANG ; Xiaofei CHEN ; Yue LIU ; Zhenyu ZHU ; Yan CAO ; Zhanying HONG ; Yifeng CHAI
Acta Pharmaceutica Sinica B 2022;12(7):3113-3123
P-glycoprotein (P-gp) highly expressed in cancer cells can lead to multidrug resistance (MDR) and the combination of anti-cancer drugs with P-gp inhibitor has been a promising strategy to reverse MDR in cancer treatment. In this study, we established a label-free and detergent-free system combining surface plasmon resonance (SPR) biosensor with styrene maleic acid (SMA) polymer membrane proteins (MPs) stabilization technology to screen potential P-gp inhibitors. First, P-gp was extracted from MCF-7/ADR cells using SMA polymer to form SMA liposomes (SMALPs). Following that, SMALPs were immobilized on an SPR biosensor chip to establish a P-gp inhibitor screening system, and the affinity between P-gp and small molecule ligand was determined. The methodological investigation proved that the screening system had good specificity and stability. Nine P-gp ligands were screened out from 50 natural products, and their affinity constants with P-gp were also determined. The in vitro cell verification experiments demonstrated that tetrandrine, fangchinoline, praeruptorin B, neobaicalein, and icariin could significantly increase the sensitivity of MCF-7/ADR cells to Adriamycin (Adr). Moreover, tetrandrine, praeruptorin B, and neobaicalein could reverse MDR in MCF-7/ADR cells by inhibiting the function of P-gp. This is the first time that SMALPs-based stabilization strategy was applied to SPR analysis system. SMA polymer can retain P-gp in the environment of natural lipid bilayer and thus maintain the correct conformation and physiological functions of P-gp. The developed system can quickly and accurately screen small molecule ligands of complex MPs and obtain affinity between complex MPs and small molecule ligands without protein purification.
7.UHPLC-Q-TOF/MS analysis of chemical constituents in compound Jinqiancao granules
Zhe ZHANG ; Zhuoran SUN ; Pengchao PAN ; Xiaofei CHEN ; Yifeng CHAI
Journal of Pharmaceutical Practice 2022;40(2):146-151
Objective To qualitatively analyze the main chemical components in compound Jinqiancao granules by ultra high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS). Methods XBridge BEH C18 column (2.1 mm×100 mm, 2.5 µm) was used for chromatographic separation. The mobile phase was composed of 0.1% formic acid water and 0.1% formic acid-acetonitrile, gradient elution, and the flow rate was 0.4 ml/min. Mass spectrometry was characterized by Quadrupole time-of-flight mass spectrometry (Q-TOF/MS) and positive ion mode scanning. Results Under the optimized LC/MS condition, 47 components in compound Jinqiancao granules were identified. The isomers were distinguished by software calculation. The source of medicinal materials was assigned. Conclusion A rapid and efficient analytical method was established for the identification of chemical components in compound Jinqiancao granules by UHPLC-Q-TOF/MS.
8.Advances in methodologies for preparation and analysis of new biochromatic stationary phase
Xinyi CHAI ; Yanqiu GU ; Xiaofei CHEN ; Yifeng CHAI
Journal of Pharmaceutical Practice 2022;40(3):193-198
Biochromatography is a new chromatographic technology with great development potential. It has been widely used in drug screening and biomolecular interaction analysis. The core of this technology is the chromatographic stationary phase of biomolecules. Nowadays, it mainly develops cell membrane chromatography, artificial biomimetic membrane chromatography and the various immobilization strategies to directly immobilizes proteins on the stationary phase carrier. This paper reviews the research progress of new biochromatographic stationary phase and the application of biochromatographic analysis based on new stationary phase. And, the applications of biochromatographic stationary phase and micro biochromatographic analysis system based on monolithic column are prospected.
9.Screening of immune cell activators from Astragali Radix using a comprehensive two-dimensional NK-92MI cell membrane chromatography/C18 column/time-of-flight mass spectrometry system
Xinyi CHAI ; Yanqiu GU ; Lei LV ; Chun CHEN ; Fei FENG ; Yan CAO ; Yue LIU ; Zhenyu ZHU ; Zhanying HONG ; Yifeng CHAI ; Xiaofei CHEN
Journal of Pharmaceutical Analysis 2022;12(5):725-732
Astragali Radix(AR)is a clinically used herbal medicine with multiple immunomodulatory activities that can strengthen the activity and cytotoxicity of natural killer(NK)cells.However,owing to the complexity of its composition,the specific active ingredients in AR that act on NK cells are not clear yet.Cell membrane chromatography(CMC)is mainly used to screen the active ingredients in a complex system of herbal medicines.In this study,a new comprehensive two-dimensional(2D)NK-92MI CMC/C18 column/time-of-flight mass spectrometry(TOFMS)system was established to screen for potential NK cell acti-vators.To obtain a higher column efficiency,3-mercaptopropyltrimethoxysilane-modified silica was synthesized to prepare the NK-92MI CMC column.In total,nine components in AR were screened from this system,which could be washed out from the NK-92MI/CMC column after 10 min,and they showed good affinity for NK-92MI/CMC column.Two representative active compounds of AR,isoastragaloside Ⅰ and astragaloside Ⅳ,promoted the killing effect of NK cells on K562 cells in a dose-dependent manner.It can thus suggest that isoastragaloside Ⅰ and astragaloside Ⅳ are the main immunomodulatory compo-nents of AR.This comprehensive 2D NK-92MI CMC analytical system is a practical method for screening immune cell activators from other herbal medicines with immunomodulatory effects.
10.Development of a surface plasmon resonance biosensor for accurate and sensitive quantitation of small molecules in blood samples
Minyu QI ; Diya LV ; Ying ZHANG ; Dongyao WANG ; Xiaofei CHEN ; Zhenyu ZHU ; Zhanying HONG ; Yifeng CHAI ; Hai ZHANG ; Yan CAO
Journal of Pharmaceutical Analysis 2022;12(6):929-936
Therapeutic drug monitoring(TDM)has played an important role in clinical medicine for precise dosing.Currently,chromatographic technology and immunoassay detection are widely used in TDM and have met most of the needs of clinical drug therapy.However,some problems still exist in practical appli-cations,such as complicated operation and the influence of endogenous substances.Surface plasmon resonance(SPR)has been applied to detect the concentrations of small molecules,including pesticide residues in crops and antibiotics in milk,which indicates its potential for in vivo drug detection.In this study,a new SPR-based biosensor for detecting chloramphenicol(CAP)in blood samples was developed and validated using methodological verification,including precision,accuracy,matrix effect,and extraction recovery rate,and compared with the classic ultra-performance liquid chromatography-ultraviolet(UPLC-UV)method.The detection range of SPR was 0.1-50 ng/mL and the limit of detec-tion was 0.099±0.023 ng/mL,which was lower than that of UPLC-UV.The intra-day and inter-day ac-curacies of SPR were 98%-114%and 110%-122%,which met the analysis requirement.The results show that the SPR biosensor is identical to UPLC-UV in the detection of CAP in rat blood samples;moreover,the SPR biosensor has better sensitivity.Therefore,the present study shows that SPR technology can be used for the detection of small molecules in the blood samples and has the potential to become a method for therapeutic drug monitoring.

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