1.Quality Evaluation of Lycii Cortex and Roasted Lycii Cortex Based on Fingerprint and Content Determination
Yihuan WU ; Wenli ZENG ; Xuemei QIN ; Zongxin SHI ; Chengcheng HUANG ; Yuntao DAI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):165-172
ObjectiveTo establish fingerprint profiles and a quantitative determination method for Lycii Cortex, providing a scientific basis for the formulation of quality standards for Lycii Cortex and its roasted products. MethodsHigh performance liquid chromatography(HPLC) was developed for the quantitative method for determining kukoamine B in Lycii Cortex and its roasted products on an Alphasil XD-C18 CH column(4.6 mm×250 mm, 5 μm). HPLC fingerprint profiles were established for 10 batches of Lycii Cortex and its roasted products, and ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to identify the common peaks based on reference standards, literature and MS information. Quality evaluation indicators included yield of decoction pieces, appearance properties, content of kukoamine B, and fingerprint profiles. The temperature and time of the roasting process were investigated to select the optimal preparation process, which was then verified. Additionally, chemical pattern recognition was combined to assess the differences in the chemical composition of Lycii Cortex before and after roasting, as well as among samples from different origins. ResultsQuantitative analysis indicated that the contents of kukoamine B in Lycii Cortex and its roasted products were 0.35%-5.51% and 0.24%-4.15%, respectively. The transfer rate of kukoamine B was 58.6%-78.9% after roasting. The fingerprint profile analysis demonstrated that the method established in this study effectively separated kukoamine B from other components in the samples and distinctly differentiated it from its impurity peak, cis-N-caffeoylputrescine. The HPLC fingerprint profiles of Lycii Cortex and its roasted products showed high similarity(all above 0.95), with 7 common peaks identified and five common components, including kukoamine B, cis-N-caffeoylputrescine, N-coumaroyl tyramine, feruloyltyramine, and glucosyringic acid, confirmed. Process optimization confirmed that baking at 110 ℃ for 20 min was a stable and feasible method for roasting Lycii Cortex. Principal component analysis and cluster analysis showed that there was little difference in the chemical composition between raw and roasted Lycii Cortex, but the quality of Lycii Cortex from different origins differed greatly. ConclusionThis study successfully established the fingerprint profiles and a quantitative method for the effective component kukoamine B in Lycii Cortex and roasted Lycii Cortex. The qualitative and quantitative analyses clarified that the impact of the roasting process on the chemical composition of Lycii Cortex was less significant than the variations due to its geographical origin. The findings of this study offer a reference for the development of quality evaluation methods and the establishment of quality standards for Lycii Cortex and its processed products.
2.Effects of Jianpi Yishen Huatan Formula (健脾益肾化痰方)-Medicated Serum on Apoptosis,Migration and the LncRNA ALAL-1/USP4/HDAC2 Pathway in Human Lung Squamous Carcinoma Cells
Yijun FANG ; Xuemei WANG ; Changzhou XIONG ; Liubang LI ; Huimin QIN ; Zhiguang WANG
Journal of Traditional Chinese Medicine 2025;66(14):1481-1488
ObjectiveTo investigate the potential mechanism of the Jianpi Yishen Huatan Formula (健脾益肾化痰方,JPYSHF) in treating squamous cell lung cancer through the LncRNA ALAL-1/USP4/HDAC2 signaling pathway. MethodsForty Sprague-Dawley (SD) rats were randomly divided into a control group and high-, medium-, and low-dose JPYSHF group with 10 rats in each group. Rats in the JPYSHF groups were administered JPYSHF concentrated liquid at doses of 45, 30, and 15 g/(kg·d) via intragastric gavage, respectively, while the control group received 10 ml/(kg·d) of normal saline, once daily for 10 consecutive days before preparation of drug containing serum. Human lung squamous carcinoma SK-MES-1 cells were divided into a control group and low-, medium-, and high-dose JPYSHF-medicated serum groups. The control group was cultured with 10% saline-containing serum, while the JPYSHF groups were cultured with 10% low-, medium-, or high-dose medicated serum. After 48 hours of incubation, flow cytometry was used to detect apoptosis rates, and a cell scratch assay was performed to evaluate migration areas at 0 h and 24 h to calculate migration rate. Additional SK-MES-1 cells were divided into control serum, JPYSHF-medicated serum (low-, medium-, high-) dose, LncRNA-silenced group (transfected with ALAL-1 siRNA), USP4-inhibited group (treated with 35 μmol/L PR-619, a deubiquitinase inhibitor), and HDAC2-inhibited group (treated with 60 μmol/L Vorinostat). After 24 and 48 hours of culture, cell viability was assessed using the CCK-8 assay; LncRNA ALAL-1, USP4, and HDAC2 mRNA levels were quantified by qPCR after 24 hours; USP4 and HDAC2 protein levels were measured by Western Blot after 48 hours. ResultsCompared with the control serum group, the total apoptosis rate of cells in middle- and high-JPYSHF-medicated serum group significantly increased, and the cell migration rate of cells in the low-, middle- and high-JPYSHF-medicated serum group significantly decreased (P<0.05 or P<0.01). The cell migration rate of the low-, medium- and high-JPYSHF-medicated serum groups decreased with the increase of concentration in a concentration-dependent manner (P<0.05 or P<0.01). Compared with the control serum group at the same time, the cell viability at 24 h and 48 h significantly decreased in all groups (P<0.05 or P<0.01). Compared with the low-JPYSHF-medicated serum group at the same time, the cell viability at 24 h and 48 h also decreased in the high-JPYSHF-medicated serum group and the LncRNA silencing group (P<0.05). Compared with the control serum group, the expression of USP4 and HDAC2 mRNA reduced in the low- and medium-dose JPYSHF-medicated serum groups and the USP4 inhibitor group, and the expression of LncRNA ALAL-1, USP4 and HDAC2 mRNA reduced in the high-dose JPYSHF-medicated serum group and LncRNA-silencing group, and HDAC2 mRNA expression reduced in the HDAC2 inhibitor group. USP4 and HDAC2 protein levels were reduced in cells of all groups except for USP4 protein level in HDAC2 inhibitor group (P<0.05 or P<0.01). ConclusionJPYSHF-medicated serum inhibits proliferation and promotes apoptosis of human lung squamous carcinoma cells, and its mechanism of action may be related to its inhibition of the LncRNA ALAL-1/USP4/HDAC2 pathway, with best effect at a high concentration.
3.Riboflavin metabolism involved into comparison of therapeutic action of wild-simulated and transplanted Astragali Radix to treat chronic atrophic gastritis rats based on endogenous-xenobiotics metabolism.
Kaidi FAN ; Xu LIAN ; Weicong ZHANG ; Xuemei QIN ; Yuetao LIU
Chinese Herbal Medicines 2025;17(3):513-528
OBJECTIVE:
Astragali Radix (AR, Huangqi in Chinese) has been utilized generally as a bulk drug for the treatment of chronic atrophic gastritis (CAG) in China. The efficacy of wild-simulated AR (WAR) and transplanted AR (TAR) commercially available is unclear. This study aimed to further clarify the therapeutic action of WAR and TAR to treat CAG rats based on endogenous-xenobiotics metabolism.
METHODS:
Ultra-high performance liquid chromatography coupled with quadrupole-time of flight mass spectrometer (UHPLC-Q-TOF/MS) based metabolomics had been used to analyze the cecal contents metabolic features and metabolic process differences of two ARs in the treatment of CAG.
RESULTS:
Apparent pharmacodynamic indicator examination revealed that the WAR group had a more substantial curative effect. Metabolomics studies revealed that seven endogenous metabolites were significant differences in two ARs. Furthermore, following treatment, 77 and 65 xenobiotics metabolites were identified in the WAR and TAR groups, respectively. Finally, the most critical riboflavin metabolic route in the formation of CAG was chosen for molecular docking with the relevant exogenous components, and WAR scored higher than TAR.
CONCLUSION
In this work, we analyzed the efficacy differences of AR from diverse cultivation forms by combining metabolomics and medicinal chemistry technologies, and it gave a fresh perspective for TCM efficacy evaluation and quality control research.
4.Relationship between tumor-infiltrating lymphocytes,PD-1 and PD-L1 expression with the prognosis of peritoneal mesothelioma
Yue CHEN ; Xuemei DU ; Zhonghe JI ; Ying GAO ; Zhiran YANG ; Huamin QIN
Chinese Journal of Clinical and Experimental Pathology 2024;40(8):828-833
Purpose To investigate the relationship be-tween the expressions of CD3,CD4,CD8,CD20,CD68,PD-1 and PD-L1 and the clinical prognosis of peritoneal mesothelioma(PM).Methods Clinical data of 69 PM patients were collect-ed.EnVision two-step immunohistochemical method was used to detect the expression of CD3,CD4,CD8,CD20,CD68,PD-1 and PD-L1 in PM.Associations between expression levels and survival were estimated by univariate and multivariate Cox pro-portional-hazards models.Results There were no significant differences in the expressions of CD3,CD4,CD8,CD20,CD68,and PD-1 in tumor infiltrating lymphocytes(TILs)of ep-ithelioid and non-epithelioid PM.The expression of PD-L1 in non-epithelioid PM TILs was higher than that in epithelioid PM TILs,but the difference was not statistically significant.The median overall survival(mOS)time of PM was 19.1 months.Multivariate models identified asbestos exposure(P=0.002),PCI score(P=0.034),histological type(P=0.036),and CD4 expression(P=0.043)was independent prognostic factors for PM.Conclusion Asbestos exposure,PCI score,histologi-cal type and CD4 expression in TILs may exert significant im-pacts on survival of PM patients.
5.Construction and application effect evaluation of ointment massage for postoperative rehabilitation of newborn after intestinal surgery
Jing RUAN ; Jiaxuan MAI ; Linghua CHEN ; Lijiao QIN ; Ling CHEN ; Jiaying CHEN ; Xuemei ZHONG ; Kun DAI
Chinese Journal of Practical Nursing 2024;40(14):1058-1065
Objective:To explore the effect of ointment massage on the postoperative recovery of newborns who had intestinal tract operation, aim to construct an effective post-operative ointment and massage scheme and to enrich the content of enhanced recovery after surgery in the field of newborns.Methods:Delphi expert inquiry method was used to construct the ointment message scheme. A total of 64 newborns undergone intestinal surgery from May 2022 to May 2023 in Guangdong Women and Children Hospital were selected as study subjects and divided into routine group (34 cases) and ointment group (30 cases) by random number table method. In addition to routine treatment and nursing, the newborns in the ointment group also received ointment massage. The postoperative gastrointestinal drainage volume, fasting days, the first feeding time, hospital stay and the number of cases of vomiting and abdominal distension between the two groups were compared.Results:A total of 16 experts with (38.93 ± 5.13) years old were inquired, 6 males and 10 females. The effective questionnaire response rates for the two rounds of expert inquiry were 16/16 and 15/16, the expert authority coefficients were both≥0.80, the Kendall′s W coefficients were 0.117 and 0.107, with statistically significant differences (both P<0.05). A ointment message scheme consisting of 8 acupoints and 7 kinds of traditional Chinese medicines was determined. In the routine group, there were 19 males and 15 females with (268.47 ± 11.27) gestational age. In the ointment group, there were 20 males and 10 femles with (271.93 ± 10.36) gestational age. The postoperative gastrointestinal drainage volume, fasting days, the first feeding time and the hospital stay in the ointment group were 74.0 ml, 6.0 d, 158.5 h and 13.0 d, which had significant differences with the 184.5 ml, 7.5 d, 192.0 h, 16.5 d in the routine group ( Z values were 2.07-2.45, all P<0.05). The incidence of vomiting and abdominal distension in the ointment group were both 8 cases, which were lower than the 19 and 18 cases in the routine group with a statistically significant difference ( χ2 = 5.58, 4.56, both P<0.05). Conclusions:The ointment massage in this study has a positive impact on the recovery of newborn infants after intestinal surgery, with high safety, in line with enhanced recovery after surgery concept, and is worthy of promotion.
6.Extraction process optimization for preparation of Ziyin Qinggu Pills by overall desirability and response surface methodology
Runliang YIN ; Xuemei WANG ; Xixiang LI ; Qin SU ; Jianliang WANG
International Journal of Traditional Chinese Medicine 2024;46(5):631-636
Objective:To optimize the extraction process of Ziyin Qinggu Pills.Methods:The solvent amount, extraction time and soaking time were taken as the influencing factors, and the overall desirability (OD) of the contents of timosaponin BⅡ, paeoniflorin, calycosin 7-O-glucoside, liquiritin and the dry extract yield were taken as the evaluation indexes, so that the optimal extraction process of Ziyin Qinggu Pills was optimally selected by the centralized composite design-response surface method.Results:The optimal extraction process of Ziyin Qinggu Pills was as follows: extraction time of 115 min, soaking time of 26 min, solvent volume of 10 times, and extraction twice.Conclusion:The optimal extraction process is reasonable, stable and reliable, which can provide the basis and foundation for the extraction process and further molding process of Ziyin Qinggu Pills.
7.Determination and Mechanism Prediction of Potential Active Ingredients in Erdongtang Based on UHPLC-QqQ-MS and Network Pharmacology
Xueyuan WANG ; Baoxin LI ; Shougang SHI ; Zhengjun HUANG ; Yuntao DAI ; Xuemei QIN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):28-34
ObjectiveTo establish an ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry(UHPLC-QqQ-MS) for determination of the active ingredients in Erdongtang, and to predict the targets and pathways of anti-insulin resistance action of this formula. MethodThe analysis was performed on an ACQUITY UPLC BEH C18 column(2.1 mm×100 mm, 1.7 μm) with the mobile phase of 0.1% formic acid aqueous solution(A)-acetonitrile(B) for gradient elution(0-3 min, 90%-87%A; 3-6 min, 87%-86%A; 6-9 min, 86%-83%A; 9-11 min, 83%-75%A; 11-18 min, 75%-70%A; 18-19 min, 70%-52%A; 19-22 min, 52%A; 22-25 min, 52%-5%A; 25-27 min, 5%-90%A; 27-30 min, 90%A). The contents of active ingredients in Erdongtang was detected by electrospray ionization(ESI) and multiple reaction monitoring(MRM) mode under positive and negative ion modes. On this basis, network pharmacology was applied to predict the targets and pathways of Erdongtang exerting anti-insulin resistance effect. ResultThe 20 active ingredients in Erdongtang showed good linear relationships within a certain mass concentration range, and the precision, stability, repeatability and recovery rate were good. The results of determination showed that the ingredients with high content in 15 batches of samples were baicalein(1 259.39-1 635.78 mg·L-1), baicalin(1 078.37-1 411.52 mg·L-1), the ingredients with medium content were mangiferin(148.59-217.04 mg·L-1), timosaponin BⅡ(245.10-604.89 mg·L-1), quercetin-3-O-glucuronide(89.30-423.26 mg·L-1), rutin(46.91-1 553.61 mg·L-1), glycyrrhizic acid(55.97-391.47 mg·L-1), neomangiferin(37.45-127.03 mg·L-1), nuciferine(0.89-63.48 mg·L-1), hyperoside(6.96-136.78 mg·L-1), liquiritin(30.89-122.78 mg·L-1), liquiritigenin(26.64-110.67 mg·L-1), protodioscin(58.57-284.26 mg·L-1), the ingredients with low content were wogonin(7.16-20.74 mg·L-1), pseudoprotodioscin(5.49-22.96 mg·L-1), ginsenoside Rb1(7.31-23.87 mg·L-1), ginsenoside Rg1(10.78-28.33 mg·L-1), ginsenoside Re(7.78-24.76 mg·L-1), ophiopogonin D(2.08-4.29 mg·L-1), methylophiopogonanone A(0.74-1.67 mg·L-1). The results of network pharmacology indicated that the mechanism of anti-insulin resistance exerted by Erdongtang might be related to the phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt) signaling pathway. ConclusionThe established UHPLC-QqQ-MS has the advantages of simple sample processing, strong exclusivity and high sensitivity, and can simultaneously determine the contents of the main ingredients from seven herbs in Erdongtang, which can lay the foundation for the development of Erdongtang compound preparations. The results of the network pharmacology can provide a reference for the mechanism study of Erdongtang in the treatment of type 2 diabetes mellitus.
8.Effects of Xiaoyao San on exercise capacity and liver mitochondrial metabolomics in rat depression model.
Weidi ZHAO ; Cui JI ; Jie ZHENG ; Shi ZHOU ; Junsheng TIAN ; Yumei HAN ; Xuemei QIN
Chinese Herbal Medicines 2024;16(1):132-142
OBJECTIVE:
This study aimed to investigate the therapeutic effects of Xiaoyao San (XYS), a herbal medicine formula, on exercise capacity and liver mitochondrial metabolomics in a rat model of depression induced by chronic unpredictable mild stress (CUMS).
METHODS:
A total of 24 male SD rats were randomly divided into four groups: control group (C), CUMS control group (M), Venlafaxine positive treatment group (V), and XYS treatment group (X). Depressive behaviour and exercise capacity of rats were assessed by body weight, sugar-water preference test, open field test, pole test, and rotarod test. The liver mitochondria metabolomics were analyzed by using liquid chromatography-mass spectrometry (LC-MS) method. TCMSP database and GeneCards database were used to screen XYS for potential targets for depression, and GO and KEGG enrichment analyses were performed.
RESULTS:
Compared with C group, rats in M group showed significantly lower body weight, sugar water preference rate, number of crossing and rearing in the open field test, climbing down time in the pole test, and retention time on the rotarod test (P < 0.01). The above behaviors and exercise capacity indices were significantly modulated in rats in V and X groups compared with M group (P < 0.05, 0.01). Compared with C group, a total of 18 different metabolites were changed in the liver mitochondria of rats in M group. Nine different metabolites and six metabolic pathways were regulated in the liver mitochondria of rats in X group compared with M group. The results of network pharmacology showed that 88 intersecting targets for depression and XYS were obtained, among which 15 key targets such as IL-1β, IL-6, and TNF were predicted to be the main differential targets for the treatment of depression. Additionally, a total of 1 553 GO signaling pathways and 181 KEGG signaling pathways were identified, and the main biological pathways were AGE-RAGE signaling pathway, HIF-1 signaling pathway, and calcium signaling pathway.
CONCLUSION
XYS treatment could improve depressive symptoms, enhance exercise capacity, positively regulate the changes of mitochondrial metabolites and improve energy metabolism in the liver of depressed rats. These findings suggest that XYS exerts antidepressant effects through multi-target and multi-pathway.
9.Clinical characteristics,effectiveness and cost of different treatment methods for invasive Klebsiella pneumoniae liver abscess syndrome
Qin HU ; Jingying LU ; Binbin DENG ; Xuemei TANG ; Zhouhua HOU
Journal of Central South University(Medical Sciences) 2024;49(5):748-757
Objective:Bacterial liver abscess is one of the common infectious diseases of the digestive system.Invasive Klebsiella pneumoniae liver abscess syndrome(IKLAS)refers to cases where,in addition to liver abscess,there are migratory infections foci or other invasive manifestations.The clinical characteristics and risk factors of IKLAS are not fully elucidated,and there is a lack of research on the effectiveness and cost-effectiveness of different treatment methods.This study aims to compare the clinical characteristics of patients with IKLAS and non-IKLAS,and explore effective and economical treatment methods. Methods:This retrospective study collected medical records of patients with Klebsiella pneumoniae liver abscess treated at Xiangya Hospital of Central South University from January 2010 to December 2023.A total of 201 patients were included,dividing into an IKLAS group(n=37)and a non-IKLAS group(n=164).Differences in demographics,symptoms and signs,laboratory indicators,imaging characteristics,comorbidities,treatment methods,treatment outcomes,and direct treatment costs between 2 groups were analyzed.The study also compared the effectiveness and costs of different treatment methods. Results:Compared with the non-IKLAS group,the proportion of patients with diabetes,Quick Sequential Organ Failure Assessment(qSOFA)≥2,immune deficiency,anemia,and thrombocytopenia in the IKLAS group was higher,and the level of procalcitonin at the onset in the IKLAS group was also higher(all P<0.05).In terms of symptoms and signs,the IKLAS group had a higher proportion of visual abnormalities and a lower proportion of complaints of abdominal pain(both P<0.05).In terms of complications,the incidence of combined pleural effusion,pulmonary infection,acute renal failure,respiratory failure,and multiple organ failure was higher in the IKLAS group(all P<0.05).The IKLAS group had a higher proportion of patients treated with antibiotics alone(24.32%vs 11.59%),while the non-IKLAS group had a higher proportion of patients treated with antibiotics combined with puncture and drainage(86.59%vs 64.86%,both P<0.05).The overall effective rate of the IKLAS group(83.78%)was lower than that of the non-IKLAS group(95.73%),and the treatment and drug costs were higher(all P<0.05).The treatment method of antibiotics combined with surgical resection of infectious foci showed a 100%improvement rate,antibiotics combined with abscess puncture and drainage had an 84.9%improvement rate,and in antibiotics alone had an 82.1%improvement rate,with statistical differences among the 3 treatment methods(P<0.05).In terms of treatment costs,antibiotics alone were the most expensive(P<0.05). Conclusion:Patients with IKLAS have poorer prognosis and higher direct medical costs.The combination of abscess puncture and drainage or surgery has a higher improvement rate and lower hospitalization costs compared to antibiotics alone,suggesting that surgical intervention may reduce antibiotic costs and save medical expenses.
10.Effect of radix scutellariae microemulsion gel on chronic eczema in mice model
Ke WANG ; Chenbing NIU ; Yinxia CHANG ; Xuemei QIN ; Zhihua LUAN ; Yue ZHANG
Chinese Journal of Clinical Pharmacology and Therapeutics 2024;29(4):447-454
AIM:To prepare radix scutellariae mi-croemulsion gel and investigate its therapeutic ef-fect on chronic eczema based on the previous re-search of radix scutellariae self microemulsion.METHODS:The gel matrix and humectant were op-timized by single factor method and response sur-face method to obtain the formula and preparation technique of the gel.The Franz diffusion cell meth-od was used to evaluate the transdermal proper-ties of microemulsion and microemulsion gel in vi-tro.By establishing a chronic eczema model in the mouse ear,the swelling degree,swelling inhibition rate,pathological changes and tumor necrosis fac-tor a(TNF-a),Interleukin-1β(IL-1β)and interleukin-6(IL-6)of radix scutellariae microemulsion gel were measured,to investigate the therapeutic ef-fect on chronic eczema in mice.RESULTS:The physi-cal and chemical properties of radix scutellariae mi-croemulsion gel were stable.Compared with micro-emulsion,the microemulsion gel had better trans-dermal performance.The cumulative transdermal amount of baicalein and wogonin,the main compo-nents of microemulsion gel,was 1.85 times and 2.77 times of that of microemulsion respectively.Moreover,the steady flow rate and permeability co-efficient of microemulsion gel significantly in-creased,and the lag time significantly shortened.Pharmacodynamic study showed that compared with the model group,the radix scutellariae micro-emulsion gel could significantly reduce the ear swelling of mice(P<0.05),and the serum inflamma-tory factor TNF-a,IL-1β and IL-6 reduced content by over 37%.Compared with the radix scutellaria aqueous extract and aqueous extract gel,the treat-ment of chronic eczema was better.CONCLUSION:The preparation process of radix scutellaria micro-emulsion gel is feasible,with strong transdermal property,and a significant therapeutic effect on chronic eczema.

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