1.Analysis on Pharmacodynamic Material Basis and Mechanism of Famous Classical Formula Renshen Wuweizi Tang in Treatment of Spleen and Lung Qi Deficiency Syndrome
Shanshan LI ; Yute ZHONG ; Xiaomei XIANG ; Wei KANG ; Shufan ZHOU ; Ping WANG ; Haiyu XU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):31-39
ObjectiveBased on ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS), network pharmacology and molecular docking techniques, to explore the pharmacodynamic material basis and mechanism of Renshen Wuweizi Tang in treating spleen-lung Qi deficiency syndrome. MethodsThe chemical components in the decoction of Renshen Wuweizi Tang were systematically characterized and identified by UPLC-Q-TOF-MS/MS, and network pharmacology was used to screen potential active ingredients, collect component targets and gene sets related to spleen-lung Qi deficiency syndrome, and obtain protein interaction relationships through STRING. Cytoscape 3.9.1 was used to construct a "formula-syndrome" association network and calculate topological feature values. Gene ontology(GO) function and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis were performed on core genes to explore potential pharmacodynamic links, the average shortest path between the formula-drug target network and the pharmacodynamic link gene network was calculated to discover dominant pharmacodynamic links, and MCODE plugin was used to identify core gene clusters from the dominant pharmacodynamic links, which were validated using Gene Expression Omnibus(GEO), and molecular docking was performed between key components and core targets. ResultsOne hundred and thirty-seven components were identified in the negative ion mode, and eighty components were identified in the positive ion mode. After deduplication, a total of 185 components were identified, mainly composed of triterpenoid saponins(49) and flavonoids(54). Based on the "formula-syndrome" correlation network analysis, energy metabolism was determined to be the dominant pharmacodynamic link of Renshen Wuweizi Tang in the treatment of spleen-lung Qi deficiency syndrome. The results of molecular docking showed that 7 components(adenosine, atractylenolide Ⅱ, atractylenolide Ⅲ, ginsenoside Rg1, glycyrrhizin B2, glycyrrhizin E2 and campesterol) from 4 medicinal materials(Ginseng Radix et Rhizoma, Atractylodis Macrocephalae Rhizoma, Glycyrrhizae Radix et Rhizoma and Poria) in this formula might regulate energy metabolism by acting on 6 targets, namely cyclic adenosine monophosphate-response element binding protein 1(CREB1), glyceraldehyde-3-phosphate dehydrogenase(GAPDH), interleukin(IL)-6, nuclear transcription factor(NF)-κB1, peroxisome proliferator-activated receptor α(PPARα), and tumor necrosis factor(TNF), thus improving the symptoms of diseases related to spleen-lung Qi deficiency syndrome. ConclusionThis study established a UPLC-Q-TOF-MS/MS for rapid characterization and identification of chemical components in the decoction of Renshen Wuweizi Tang, expanding the understanding of the material composition of this formula, and found that 7 components might act on the key advantageous pharmacodynamic link "energy metabolism" through 6 targets to improve the related symptoms of spleen-lung Qi deficiency syndrome. This can provide a reference for the subsequent exploration of the material benchmark and mechanism of the famous classical formula.
2.Improvement on Quality Standard of Yuanhu Zhitong Oral Liquid
Lu FU ; Chengyu CHEN ; Jin GAO ; Dan WU ; Chun LI ; Zhiming CAO ; Jianli GUAN ; Ping WANG ; Haiyu XU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):125-131
ObjectiveTo improve the quality standard of Yuanhu Zhitong oral liquid in order to strengthen the quality control of this oral liquid. MethodThin layer chromatography(TLC) was used for the qualitative identification of Corydalis Rhizoma and Angelicae Dahuricae Radix in Yuanhu Zhitong oral liquid by taking tetrahydropalmatine, corydaline reference substances and Corydalis Rhizoma reference medicinal materials as reference, and cyclohexane-trichloromethane-methanol(5∶3∶0.5) as developing solvent, Corydalis Rhizoma was identified using GF254 glass thin layer plate under ultraviolet light(365 nm). And taking petroleum ether(60-90 ℃) -ether-formic acid(10∶10∶1) as developing solvent, Angelicae Dahuricae Radix was identified using a silica gel G TLC plate under ultraviolet light(305 nm). High performance liquid chromatography(HPLC) was performed on a Waters XSelect HSS T3 column(4.6 mm×250 mm, 5 μm) with acetonitrile(A)-0.1% glacial acetic acid solution(adjusted pH to 6.1 by triethylamine)(B) as the mobile phase for gradient elution(0-10 min, 20%-30%A; 10-25 min, 30%-40%A; 25-40 min, 40%-50%A; 40-60 min, 50%-60%A), the detection wavelength was set at 280 nm, then the fingerprint of Yuanhu Zhitong oral liquid was established, and the contents of tetrahydropalmatine and corydaline were determined. ResultIn the thin layer chromatograms, the corresponding spots of Yuanhu Zhitong oral liquid, the reference substances and reference medicinal materials were clear, with good separation and strong specificity. A total of 12 common peaks were identified in 10 batches of Yuanhu Zhitong oral liquid samples, and the peaks of berberine hydrochloride, dehydrocorydaline, glaucine, tetrahydropalmatine and corydaline. The similarities between the 10 batches of samples and the control fingerprint were all >0.90. The results of determination showed that the concentrations of corydaline and tetrahydropalmatine had good linearity with paek area in the range of 0.038 6-0.193 0, 0.034 0-0.170 0 g·L-1, respectively. The methodological investigation was qualified, and the contents of corydaline and tetrahydropalmatine in 10 batches of Yuanhu Zhitong oral liquid samples were 0.077 5-0.142 9、0.126 1-0.178 2 g·L-1, respectively. ConclusionThe established TLC, fingerprint and determination are simple, specific and reproducible, which can be used to improve the quality control standard of Yuanhu Zhitong oral liquid.
3.Quantitative MRI analysis of anterior cruciate ligament sprain and chronic injury of knee joint and comparison study with arthroscopy
Haiyu ZHANG ; Yutao YAN ; Shuo ZHANG ; Yuebin WANG
Journal of Practical Radiology 2024;40(4):609-612
Objective To study the application value of 3.0T MRI T2 mapping quantitative technology in the diagnosis of anterior cruciate ligament sprain and chronic injury of knee joint.Methods A total of 82 subjects were studied,and the experimental group 72 cases was divided into grade Ⅰ injury group(25 cases),grade Ⅱ injury group(25 cases),chronic injury group(22 cases),and control group 10 cases.The experimental group met the criteria of arthroscopy.The proximal,middle,and distal segments of the anterior cruciate ligament were selected as the region of interest(ROI),and T2 mapping values were measured.The differences in T2 mapping values of each area were compared between and within the groups,while compared with arthroscopy.Results The T2 mapping values in grade Ⅰ,Ⅱ,and chronic injury groups were higher than those in control group(P<0.05).Comparison within the experimental group:the T2 mapping values of each area in grade Ⅱ injury group were higher than those in grade Ⅰ injury group and chronic injury group(P<0.05).The T2 mapping values of each area in grade Ⅰ injury group were higher than those in chronic injury group(P<0.05).The specificity,sensitivity,positive predictive value,negative predictive value and accuracy of T2 mapping in diagnosing anterior cruciate ligament grade Ⅰ injury were 94.7%,95.5%,89.7%,96.6%,and 90.2%respectively.The specificity,sensitivity,positive predictive value,negative predictive value,and accuracy of grade Ⅱ injury were 89.4%,87.9%,92.1%,93.4%,and 93.8%respectively.The specificity,sensitivity,positive predictive value,negative predictive value,and accuracy of chronic injury were 92.2%,95.4%,90.3%,87.6%,and 91.5%respectively.Kappa test showed a good con-sistency between T2 mapping results and arthroscopic results,with a Kappa value of 0.763(P<0.01).Conclusion The value of MRI T2 mapping can provide a reference for the clinical diagnosis of anterior cruciate ligament sprain and chronic injury of knee joint,and the results are in good agreement with the control of arthroscopy.
4.Development and evaluation of a rapid and sensitive POC chemiluminescent assay for β-human chorionic gonadotropin
Haiyu XIE ; Jing QIN ; Yanni ZHANG ; Junjie LIU ; Xiaowei HE ; Yu WANG
Chinese Journal of Immunology 2024;40(3):615-620
Objective:To develop and evaluate a rapid and sensitive point-of-care chemiluminescent assay(POC-CLIA)for β-human chorionic gonadotropin(β-HCG).Methods:POC-CLIA was constructed based on alkaline phosphatase(Alp)-AMPPD lumi-nescence system and magnetic particles(Mps)carrier.Performance of POC-CLIA,including sensitivity,precision,accuracy,linear dilution,specificity,stability,hook effect and clinical application were evaluated.Results:Detection limit of β-HCG was 0.71 mU/ml,linear detection range was 0.710~1.092×104 mU/ml,and was no hook effect up to 1.7×105 mU/ml.Intra and inter batch coefficients of variation were less than 10%,and could be stored stably at 37℃ for 10 days.Accuracy deviation was within±10%,so results were reliable.There was no cross-reactivity between interfering substances and anti-β-HCG antibdies.For detecting β-HCG in 100 clinical serum samples,results were highly correlated with those that were tested by clinical standard methods(R2=0.997 0).Turnaround time for single sample was less than 15 min and throughput could reach 200 T/h.Conclusion:This method is adequate that can be widely used in grassroots communities to help large-scale screening of pregnancy and related diseases.
5.Clinical application and efficacy analysis of castor stent graft in the treatment of aortic diseases
Jiaxiang ZHUANG ; Ren WANG ; Xianlu MA ; Qi XIE ; Zhi DOU ; Fuzhen ZHENG ; Haiyu CHEN ; Yuanxiang CHEN ; Licheng YAN ; Hongjie ZHOU
Chinese Journal of Thoracic and Cardiovascular Surgery 2024;40(2):79-84
Objective:To summarize the use of Castor stent graft in aortic diseases and to analyze their efficacy.Methods:The clinical data of patients with aortic diseases treated with Castor stent graft from November 2017 to August 2022 in Fujian Provincial Hospital were collected and divided into branched stent group and branched stent co-operative group according to the operation method, and the clinical data of both groups were summarized.Results:A total of 75 cases of aortic disease were treated with Castor stents, and finally 53 cases were classified as branched stent group and 22 cases as branched stent co-operative group. The operations in both groups were successful. The median operative time in the branched stent group was 120(100, 160)min, and the median postoperative hospital stay was 7.0(5.5, 10.5)days.There was no postoperative ischemic stroke, no spinal cord ischemia. One case of new aortic dissection occurred. During follow-up, there was one lost case and two deaths, and the rest did not have endoleak, branch stent stenosis, ischemic stroke or re-intervention. In the branched stent co-operative group, there was one postoperative ischemic stroke, one case of slight stenosis of the left subclavian artery stent during follow-up, the remaining cases had satisfactory postoperative outcomes.Conclusion:Castor stent graft is a safe and effective procedure in the treatment of aortic diseases. And Castor stent graft can be used in combination with other endovascular repair techniques in the treatment of complex aortic diseases, with safe and reliable postoperative outcomes.
6.Analysis of Material Basis of Famous Classical Formula Baoyuantang Based on UPLC-Q-TOF-MS/MS
Wenjing GAO ; Shanshan LI ; Xiaomei XIANG ; Yi SUN ; Yang QU ; Chunling ZHOU ; Shufan ZHOU ; Lun YU ; Bing LI ; Ping WANG ; Haiyu XU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(22):243-248
ObjectiveA rapid method for identification of chemical constituents in Baoyuantang reference sample was established in order to clarify the material basis of this formula. MethodBased on ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) and self-established database information, the chemical components in Baoyuantang were systematically characterized and identified. The chromatography was performed on a Waters ACQUITY UPLC HSS T3 column(2.1 mm×100 mm, 1.8 μm) with mobile phase of 0.1% formic acid aqueous solution(A)-0.1% formic acid acetonitrile solution(B) for gradient elution(0-3 min, 2%-19%B; 3-8 min, 19%B; 8-8.1 min, 19%-22%B; 8.1-14 min, 22%-29%B; 14-16 min, 29%B; 16-32 min, 29%-45%B; 32-32.1 min, 45%-90%B; 32.1-35 min, 90%-95%B; 35-36 min, 95%-98%B; 36-37 min, 98%-2%B; 37-40 min, 2%B). Based on electrospray ionization(ESI), continuum data format was collected in both positive and negative ion modes with a scanning range of m/z 50-1 500. Chemical constituents in the decoction of Baoyuantang were systematically analyzed by UNIFI 1.9.4 software matching, control comparison, The Encyclopedia of Traditional Chinese Medicine(ETCM) database search and literature reports. ResultA total of 229 components were identified under negative ion mode and 181 under positive ion mode, with a total of 322 components after removing duplicates, including 116 triterpene saponins, 66 flavonoids, 19 organic acids, 6 gingerphenols, 6 gingerols, 5 gingerones, 10 amino acids, 7 saccharides, 5 coumarins and 82 other components. Among them, 83, 141, 39, 35 and 38 components were attributed to Ginseng Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma, Astragali Radix, Cinnamomi Cortex and Zingiberis Rhizoma Recens, respectively. ConclusionIn this study, the rapid characterization and identification of multi-class components in Baoyuantang was realized, and it was confirmed that the material basis of this formula was mainly triterpenoid saponins, flavonoids, gingerols and organic acids, and the chemical composition was attributed and analyzed, which provided a reference for the subsequent quality control research.
7.Exploring Effect of Levo-tetrahydropalmatine on Spinal Metabolic Profiles of Rats with Chronic Pain Based on Widely-targeted Metabolomics
Dan WU ; Junhong ZHANG ; Lu FU ; Yute ZHONG ; Ping WANG ; Haiyu XU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):187-194
ObjectiveTo elucidate the underlying mechanism of the efficacy of Levo-tetrahydropalmatine (l-THP) in alleviating chronic pain and identify the key metabolites and metabolic pathways for l-THP regulation. MethodA classical chronic constrictive injury (CCI) model was built in rats’ bodies, and the pain intensity was evaluated by detecting the mechanical withdrawal threshold. On the sixth day after surgery, oral administration of l-THP (64 mg·kg-1) and positive control drug pregabalin (Pre, 30 mg·kg-1) was performed on rats. After the last administration following consecutive five times of administration, ipsilateral spinal cord tissues were collected for widely-targeted metabonomics, with eight rats in each group. Differential metabolites (DEMs) were identified according to the standard of VIP>1.0 and P<0.05, and functional enrichment and interaction analyses of the Kyoto Encyclopedia of Genes and Genomes (KEGG) were performed to obtain the key metabolites and metabolic pathways associated with the analgesic effects of l-THP. ResultIn behavioral science, administration of both l-THP and Pre significantly improved mechanical hyperalgesia in CCI rats (P<0.01), thus mitigating pain. Metabonomic analysis results revealed that l-THP administration corrected the aberrant metabolic profile in the spinal cord of CCI rats. Meanwhile, 53 DEMs were called back, including several classical pain biomarkers such as sphingosine-1-phosphate (S1P), cyclic adenosine monophosphate (cAMP), acetylcholine, and glutamate. Functional enrichment analysis of the DEMs indicated the involvement of metabolic pathways such as ferroptosis, autophagy, neuroactive ligand-receptor interactions, phospholipase D and cAMP-related signaling pathways, glutathione metabolism, and cofactor biosynthesis in mediating the effects of l-THP on the metabolic profile of the spinal cord. Further analyses on the relative metabolite abundance and metabolic pathways indicated that by significantly decreasing the relative levels of glutamate (P<0.01) and glycine (P<0.01) in the spinal cord, l-THP can promote the synthesis of reduced glutathione (GSH) and increase the ratio of reduced/oxidized GSH (P<0.05). Additionally, it can relieve oxidative stress in the spinal cord of CCI rats and significantly reduce the acetyl-CoA level (P<0.01) to finally inhibit ferroptosis occurrence. Conclusionl-THP may exert analgesic effects by regulating multiple metabolic pathways including GSH metabolism, ferroptosis, cofactor biosynthesis, and amino acid synthesis to correct the aberrant metabolic profile in the spinal cord of CCI rats. Ferroptosis and GSH metabolism may be the key pathways for l-THP regulation, with glutamate, glycine, glutathione, and acetyl-CoA as the key metabolites.
8.Pharmaceutical service in a case of fat embolism syndrome following postoperative fracture
Leijiao ZHANG ; Pingping WANG ; Qinqin YAN ; Haiyu HUANG ; Guoxi HUANG ; Xue WU
China Pharmacy 2024;35(22):2822-2827
OBJECTIVE To analyze the pharmaceutical service process in a fracture patient complicated by fat embolism syndrome (FES) following postoperative fracture, aiming to provide a reference for clinical treatment and pharmaceutical service for similar patients. METHODS Clinical pharmacist participated in the entire treatment process of a patient with FES following postoperative fracture. Based on the patient’s clinical manifestations and test results, literature was reviewed to assist clinical physicians in formulating the therapeutic regimen of glucocorticoids. For the drug-related adverse reactions of renal function impairment and reduced platelet count that occurred during the treatment, suspicious drugs were analyzed and disposed of accordingly. RESULTS The clinical pharmacist recommended Hydrocortisone sodium succinate for injection (100 mg, q8 h, ivgtt, for about one week followed by a gradual dose reduction) for treating FES. The Vancomycin hydrochloride for injection used in this case was assessed as “very probably” associated with the adverse drug reactions of renal function impairment and thrombocytopenia. The clinical physician adopted the pharmacist’s medication recommendations, and the patient’s condition stabilized after treatment, with improvement in adverse reactions, and was discharged from the hospital. CONCLUSIONS The use of glucocorticoids in treating FES has a definite therapeutic efficacy. Clinical pharmacists should individualize the medication plan based on the patient’s pathological state and distinguish it from postoperative sepsis. Meanwhile, drug-induced adverse reactions in the kidney and blood system should be closely monitored.
9.Analysis of Specific Chromatogram of Classical Formula Qianghuo Shengshi Tang Reference Samples
Wenya GAO ; He XU ; Mingli LI ; Haiyu ZHAO ; Yanyan ZHOU ; Hongjie WANG ; Jian YANG ; Xiaolu WEI ; Zhikun FAN ; Nan SI ; Baolin BIAN
Chinese Journal of Modern Applied Pharmacy 2024;41(10):1350-1356
OBJECTIVE
To establish the specific chromatogram of Qianghuo Shengshi Tang(QHSS) reference sample, clarify the key quality attributes of QHSS, providing reference for the quality evaluation of QHSS reference sample.
METHODS
The SilGreen C18 column(4.6 mm×250 mm, 5 μm) was used. The mobile phase consisted acetonitrile and 0.2% formic acid aqueous solution. The detection wavelength was 328 nm. Established an HPLC characteristic spectrum analysis method for the reference sample of QHSS. A variety of chromatographic columns and different instruments were applied to investigate the adaptability of the system. HPLC-LTQ-Orbitrap MS was used to identify the specific peaks of the QHSS reference samples in positive ion mode.
RESULTS
There were 14 peaks in the specific chromatogram, which belonged to Notopterygii Rhizoma Et Radix, Angelicae Pubescentis Radix, Ligustici Rhizoma Et Radix, Chuanxiong Rhizome, Viticis Fructus, respectively. Ferulic acid(peak 3) was reference peak. A total of 22 compounds were identified by mass spectrometry, including coumarin and flavonoids.
CONCLUSION
The established specific chromatogram method of QHSS is simple, stable and reproducible. The material basis of QHSS reference sample is basically determined, providing a reference for the development and quality control of QHSS.
10.Prognostic Value of Radiation-Induced Lymphopenia in Patients with Unresectable Primary Hepatocellular Carcinoma Receiving Radiotherapy
Jinlong TONG ; Haiyu WANG ; Xiaoqiang TIAN ; Ying LI ; Shihui LU ; Wei YE
Cancer Research on Prevention and Treatment 2024;51(12):1007-1014
Objective To evaluate the prognostic value of radiation-induced lymphocytopenia in the survival of patients with primary hepatocellular carcinoma receiving radiation therapy. Methods The clinical data of 98 patients with unresectable primary hepatocellular carcinoma who received radiotherapy were retrospectively analyzed. The minimum absolute lymphocyte count (min ALC) was graded in accordance with CTCAE V4.0. The optimal threshold of min ALC for prognosis was calculated by using the receiver operating characteristic curve, and the correlation of min ALC with clinical characteristics and dosimetry parameters was analyzed. The Kaplan-Meier method was employed to analyze the survival of patients with different levels of min ALC. Univariate and multivariate Cox proportional regression models were applied to analyze prognostic factors. Results The baseline and min ALC of 98 patients during radiotherapy were 1.52×109/L and 0.45×109/L, respectively(P<0.001). The optimal cut-off value of min ALC for the prediction of the one-year survival rate was 0.38×109/L. GTV, the mean dose of the liver and spleen, the V5 and V10 of the liver and spleen, and the V15 of the spleen were correlated with min ALC, and the V5 of the liver was an independent predictor of min ALC. The overall survival of patients with high min ALC was higher than that of patients with low min ALC. Independent prognostic factors were min ALC≤0.38×109/L (HR=0.515, P=0.024), min ALC≥grade 3 (HR=0.576, P=0.032), tumor thrombus in the portal/vena cava, Child-Pugh grade A, increase of ≥2 points in the Child-Pugh score after radiotherapy, and received more than two other therapies. Conclusion Min ALC≤0.38×109/L and min ALC≥grade 3 have independent prognostic value in patients with unresectable hepatocellular carcinoma receiving radiotherapy.


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