1.Hepatitis E virus infection among blood donors in Hangzhou
Ziyun DU ; Wenjun DENG ; Danxiao WU ; Jinhui LIU ; Jie DONG
Chinese Journal of Blood Transfusion 2025;38(1):19-25
[Objective] To investigate the infection and characteristics of hepatitis E virus among blood donors in Hangzhou. [Methods] A total of 5 075 blood samples of blood donors from Zhejiang Provincial Blood Center from September to November 2023 were collected, including 5 037 samples with normal ALT and 38 samples with elevated ALT (>50 U/L). Enzyme-linked immunosorbent assay (ELISA) was used to detect anti-HEV IgM, anti-HEV IgG and HEV-Ag. The Fisher test and Chi-square test were used to evaluate the difference in the reactivity rates of anti-HEV IgM and anti-HEV IgG among different levels of ALT. The distribution characteristics of HEV screening in blood donors were analyzed. Univariate and multivariate logistic regression were used to analyze the susceptibility factors of anti-HEV IgM and anti-HEV IgG seropositivity, and the anti-HEV IgM-reactive blood donors were followed up by telephone. [Results] The reactivity rates of anti-HEV IgM, anti-HEV IgG and HEV-Ag in 5 075 blood samples were 0.45%, 22.98% and 0%, respectively. There was no difference in the reactivity rates of anti-HEV IgM and anti-HEV IgG among different levels of ALT (P>0.05), and the results of univariate and multivariate logistic regression analysis showed that age was a risk factor for anti-HEV IgM and anti-HEV IgG reactivity in blood donors (P<0.05), while no difference in the reactivity rates of anti-HEV IgM and anti-HEV IgG among blood donors was noticed in gender, occupation and education level (P>0.05). [Conclusion] There is a potential risk of transfusion-transmitted HEV (TT-HEV) in Hangzhou, and a cost-effective HEV screening strategy needs to be established to continue regular HEV surveillance in Hangzhou to assess the risk of infection.
2.Clinical Observation and Mechanism Study of Xu's Shenqi Yizhu Decoction Combined with Chemotherapy in the Treatment of Cancer-related Fatigue of Stagnated-toxin Spleen Deficiency Type in Postoperative Gastric Cancer Patients
Zhixiang SHEN ; Weimin LU ; Chuan SHI ; Mengwei YE ; Mengyuan LEI ; Ziyun LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):143-151
ObjectiveTo evaluate the clinical efficacy of Xu's Shenqiyizhu (SQYZ) decoction combined with chemotherapy in the treatment of cancer-related fatigue (CRF) of stagnated-toxin spleen deficiency type after gastric cancer surgery and explore its possible mechanism. MethodsFifty postoperative gastric cancer patients with CRF of stagnated-toxin spleen deficiency type were selected and randomly divided into an experimental group and a control group by using a random number table,with 25 cases in each group. The control group was treated with FLOT chemotherapy (50 mg·m-2 docetaxel (iv drip on day 1) + 85 mg·m-2 oxaliplatin (iv drip on day 1) + 200 mg·m-2 calcium folinate (iv drip on day 1) + 2 600 mg·m-2 fluorouracil (iv drip for 24 h on day 1),once every three weeks) and basic and symptomatic supportive treatment. The experimental group was treated with Xu's SQYZ decoction (decocted twice,200 mL taken orally twice a day) in addition to the treatment of the control group. One course of treatment lasted for three weeks,with a total of four courses conducted. Observation was performed on the piper fatigue scale (PFS) scores,karnofsky performance status (KPS) scores,European Organization for Research and Treatment of cancer quality of life questionnaire (EORTC QLQ-C30) scores,traditional Chinese medicine syndrome scores,and serum levels of tumor necrosis factor-α (TNF-α),interferon-γ (IFN-γ),and interleukin-6 (IL-6)detected via enzyme linked immunosorbent assay (ELISA) before and after treatment in the two groups. The safety test results before and after treatment for the two groups of patients,as well as the occurrence of adverse events during treatment, were recorded. Transcriptome sequencing data of peripheral blood samples from gastric adenocarcinoma patients and normal individuals were downloaded from the gene expression omnibus (GEO) database,and differentially expressed genes between the tumor and normal groups were identified. Differential gene enrichment analysis was made based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO). The CRF relevance scores of genes were retrieved from the GeneCards database. Results① Compared with that before treatment,the total PFS score in the experimental group was significantly reduced (P<0.05). Compared with the control group after treatment,the experimental group showed significantly reduced total PFS score (P<0.05). ② Compared with that before treatment,the KPS score in the experimental group decreased significantly (P<0.05). Compared with the control group after treatment,the experimental group exhibited a significantly decreased KPS score (P<0.05). The experimental group demonstrated significantly increased functional scores (physical function,role function,emotional function,social function,and overall health) (P<0.05) and significantly reduced symptom scores (fatigue,shortness of breath,loss of appetite,constipation,and diarrhea) of the EORTC QLQ-C30 scale after treatment compared with before treatment. Compared with the control group after treatment,the experimental group presented significantly increased functional scores (physical function,emotional function,social function,and overall health) (P<0.05) and significantly reduced symptom scores (fatigue,nausea and vomiting,shortness of breath,loss of appetite,and diarrhea) of the EORTC QLQ-C30 scale (P<0.05). Compared with those before treatment,the traditional Chinese medicine syndrome scores (eating too little and poor digestion,fatigue and weakness,postprandial bloating,abnormal bowel movements,lassitude and weakness,and total score) in the experimental group were significantly reduced (P<0.05). Compared with the control group after treatment,the experimental group had significantly reduced traditional Chinese medicine syndrome scores (eating too little and poor digestion,fatigue and weakness,nausea and vomiting,and sallow complexion) (P<0.05), which indicated better efficacy in the experimental group than in the control group (χ2=7.996,P<0.05). The serum levels of TNF-α,IL-6,and IFN-γ were significantly correlated with each other (P<0.01). Compared with those before treatment,the levels of serum cytokines TNF-α,IL-6,and IFN-γ in the experimental group were significantly reduced (P<0.05). Compared with the control group after treatment,the experimental group showed significantly reduced serum levels of cytokines TNF-α,IL-6,and IFN-γ (P<0.05). ③ There were no significant intra-group and inter-group differences in the safety test results of the two groups before and after treatment. During the treatment period,there was no significant difference in the incidence of adverse reactions between the two groups of patients. ④ Compared with the normal group,the tumor group exhibited a total of 328 significantly up-regulated genes in the peripheral blood (P<0.05),and KEGG and GO analyses showed that they were significantly enriched in signaling pathways such as TNF (P<0.05). ⑤ TNF,IL6,IFNG, and other cytokine encoding genes may be key pathogenic genes for CRF. ConclusionXu's SQYZ decoction can alleviate symptoms such as fatigue in postoperative chemotherapy patients with gastric cancer and improve their functional status and quality of life. Its mechanism may be related to improving cytokine imbalance.
3.Study on the Molecular Mechanism of Shema Zhichuan Liquid in the Treatment of Neutrophilic Asthma Based on Network Pharmacology and In Vivo Experiment
Leshen LIAN ; Xingru MENG ; Xiufang HUANG ; Jinxi ZHOU ; Yanxiao XIE ; Hailan TAO ; Ziyun JIANG ; Xiaohong LIU
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(2):247-256
Objective To investigate the molecular mechanism of Shema Zhichuan Liquid in the treatment of neutrophilic asthma(NA)based on network pharmacology and in vivo experiments.Methods(1)The TCMSP,literature search and Swiss ADME and Swiss Target Prediction databases were used to search and screen the active components and their targets of Shema Zhichuan Liquid.OMIM,GeneCards,DisGeNET and DrugBank databases were used to search and screen NA disease-related targets.The intersection of the active components and NA disease-related targets of Shema Zhichuan Liquid was obtained through the microbiology platform to obtain the potential targets of Shema Zhichuan Liquid for the treatment of NA(common targets).Cytoscape 3.8 software was used to construct the network of"Chinese medicinals-active components-potential targets".The PPI network of potential targets was established by STRING database,and the core targets were obtained by analysing the built-in Mcode plug-in.The Metascape platform was used to enrich the gene ontology(GO),Kyoto Encyclopaedia of Genes and Genomes(KEGG)pathways for the potential targets.(2)BALB/C mice were acclimatised and fed for 1 week and randomly divided into a blank group,NA model group,low-dose group(2.5 g·kg-1)and high-dose group of Shema Zhichuan Liquid(10 g·kg-1),and control group of Dexamethasone(1 mg·kg-1);the NA mouse model was replicated by intraperitoneal injection of sensitizer(OVA+CFA)and nebulized inhalation excitation.OVA/CFA(20 μg OVA+75 μg CFA,0.3 mL)was injected intraperitoneally to sensitize on days 0,7 and 14 respectively,and 5%OVA suspension was nebulized on days 21-30(8 mL each time,40 minutes each time,once a day);1 hour before nebulisation,each group was administered by gastric gavage,and the Dexamethasone control group was administered by intraperitoneal injection once a day.The pathological changes of mouse lung tissue were observed by HE staining;IL-8 content in alveolar lavage fluid was detected by ELISA;mRNA expression levels of NLRP3 and CXCR2 were detected by RT-qPCR;and p-mTOR protein expression levels was detected by immunohistochemistry.Results(1)A total of 826 active component targets and 154 NA disease-related targets were obtained,and 51 potential targets(common targets)for the treatment of NA were obtained from the intersection of the active component and the NA disease-related targets of Shema Zhichuan Liquid.Through the network analysis of"Chinese medicinals-active components-potential targets",quercetin,lignocerotoxin,kaempferol,stigmasterol,naringenin and other key active components were obtained.The PPI network analysis of potential targets yielded 29 core targets,including AKT1,IL6,TNF,EGFR,NLRP3,RELA,MIF,CXCR2,VEGFA,etc..The GO functional enrichment analysis yielded 882 biological process entries,33 cellular component entries,and 61 molecular function entries;KEGG analysis yielded 142 signaling pathways,mainly involving TNF signaling pathway,influenza A signaling pathway,Toll-like receptor pathway,MAPK signaling pathway,mTOR signaling pathway and so on.(2)Results of animal experiments:compared with the blank group,mice in the NA model group showed obvious damage to the airway mucosa,structural disorders,a large number of inflammatory cells infiltration,mucosal congestion,oedema,obvious thickening of the alveolar wall,and narrowing of the alveolar lumen;the level of the inflammatory factor IL-8 in the alveolar lavage fluid was significantly elevated(P<0.05);the mRNA expressions of NLRP3 and CXCR2 in the lung tissues of the mice were significantly up-regulated(P<0.01),and the protein expression of p-mTOR was significantly increased.Compared with the NA model group,the structural arrangement of bronchial epithelial cells in the mice in the low-and high-dose groups of Shema Zhichuan Liquid was slightly disordered,with a small amount of inflammatory cell infiltration around the airways and blood vessels,and the congestion and edema of the bronchial mucosa were significantly reduced;the mRNA expression of CXCR2 in the lung tissues of the mice was significantly down-regulated(P<0.01),and the level of expression of p-mTOR protein was significantly reduced.The IL-8 level in the vesicular lavage fluid of mice in the high-dose group was significantly reduced(P<0.05);the mRNA expression of NLRP3 in the lung tissue of mice in the low-dose group was significantly down-regulated(P<0.05).Conclusion The therapeutic effect of Shema Zhichuan Liquid on NA may be achieved through the key active components,such as quercetin,lignocerol and kaempferol,acting on the core targets,such as NLRP3 and CXCR2,and regulating the key signaling pathways,such as the TNF signaling pathway,the MAPK signaling pathway,the Toll-like signaling pathway,and the mTOR pathway.
4.Discussion on the Optimal Dose of Aspirin in the Treatment of Acute Stage of Kawasaki’s Disease
Jie MI ; Zhuo LIU ; Yuan LI ; Yang LI ; Ziyun DUAN ; Wenwen ZHANG ; Jiahua LIU
Chinese Journal of Modern Applied Pharmacy 2024;41(3):386-390
OBJECTIVE
To study the effect of different doses of aspirin on clinical efficacy in acute stage of Kawasaki’s disease, and to explore the optimal dose of aspirin.
METHODS
A total of 150 patients suffered from Kawasaki’s disease were randomly selected by hospital information system from March to May 2022 for retrospective analysis. According to different doses of aspirin, they were divided into three groups: high dose group(>50 mg·kg−1·d−1), medium dose group(30−50 mg·kg−1·d−1) and low dose group(<30 mg·kg−1·d−1). The antipyretic time, the incidence of non-response to intravenous human immunoglobulin, the improvement of laboratory indexes and prevalence of adverse drug reaction were compared among the three groups.
RESULTS
There was no significant difference in body temperature recovery among the three groups under different doses of aspirin. There was no significant difference in patients with non-response to intravenous human immunoglobulin among the three groups. Before treatment, there were no significant differences in white blood cell(WBC) count, blood platelet(PLT) count and C-reactive protein(CRP) concentration among the three groups. After treatment, the count of WBC, PLT and CRP in the three groups was significantly improved compared with that before treatment, and the difference was statistically significant(P<0.05). However, there was no significant difference in the above indexes among the three groups after treatment. There was a higher incidence of adverse reactions in children treated with medium or high dose aspirin.
CONCLUSION
Different doses of aspirin combined with intravenous human immunoglobulin have good therapeutic effect on Kawasaki’s disease, but considering the safety and economy of aspirin, low dose administration is recommended.
5.Clinical analysis of acute bilateral brachium pontis infarction
Ziyun YUAN ; Ran LIU ; Lei XIANG ; Wei YUE
Chinese Journal of Neurology 2024;57(8):859-866
Objective:To investigate the clinical manifestation, imaging, pathogenesis and prognostic factors associated with acute bilateral brachium pontis infarction.Methods:Patients with acute bilateral brachium pontis infarction who were admitted to the Tianjin Huanhu Hospital from October 2016 to October 2021 were collected for retrospective analysis. The demographic information, clinical symptoms and signs, imaging and prognosis of the patients were collected. Binary Logistic regression was used to analyze the correlation between age, gender, stroke risk factors, infarct site, vascular condition, National Institutes of Health Stroke Scale (NIHSS) score on admission, stroke progression, and 90-day prognosis of patients with acute bilateral brachium pontis infarction.Results:A total of 112 patients with acute bilateral brachium pontis infarction were included. The most common clinical features were ataxia (84.8%, 95/112), and vertigo (75.0%, 84/112); the first symptom was tinnitus/hearing loss in 23 patients (20.5%, 23/112), of whom 4 patients had sudden deafness. Among the 112 patients, isolated bilateral brachium pontis infarcts were found only in 8 cases (7.1%), whereas pontine infarcts in 90 cases (80.4%), midbrain infarcts in 22 cases (19.6%), medullary infarcts in 21 cases (18.8%), and cerebellar infarcts in 86 cases (76.8%). Of the patients who underwent vascular imaging, moderate-to-severe stenosis/occlusion of the vertebrobasilar artery system was present in 89.0% (89/100), and moderate-to-severe stenosis/occlusion of the vertebral arteries and/or basilar arteries was present bilaterally in 65.0% (65/100) of the patients. Pathogenesis was predominantly the large atherosclerotic type (79.5%, 89/112). Progressive stroke ( OR=7.765,95% CI 2.760-21.841, P<0.001) and higher NIHSS score on admission ( OR=1.196,95% CI 1.085-1.318, P<0.001) were independent risk factors for poor prognosis in patients with bilateral brachium pontis infarction. Conclusions:The first symptom in patients with acute bilateral brachium pontis infarction is dizziness, and acute vestibular and hearing disorders are the characteristic manifestations of brachium pontis infarction. Isolated bilateral brachium pontis infarction is rare in clinic, and most cases are combined with posterior circulation infarction, and the pons and cerebellum are the most likely to be involved. Pathogenesis is predominantly the large atherosclerotic type. Progressive stroke and higher NIHSS score on admission are independent risk factors for poor prognosis of the patients with the disease.
6.Reconstruction of thumb degloving injury with hallux nail flap combined with 3D printing: a report of 16 cases
Ziyun YU ; Chao LIU ; Wei DING ; Shiliang GENG ; Zhongcheng LI ; Chenglong XUE ; Guihong ZHANG ; Xuetao LIU
Chinese Journal of Microsurgery 2023;46(5):540-544
Objective:To explore the effect of hallux nail flap design assisted with 3D printing in reconstruction of thumb defect in degloving injury.Methods:From January 2020 to March 2022, 16 patients with thumb defect caused by degloving injury with intact digit flexor and extensor tendons were treated. The patients were 11 males and 5 females, aged 20 to 52 years old, with an average age of 31 years old. The injured patient's hands were firstly scanned by CT and the 3D models were created to map the profile of the tissues required for reconstruction. Photopolymer templates for the defects in thumb were then 3D printed. The printed templates were put on the great toes and accordingly the hallux nail flaps were designed and harvested. The degloving wound of thumb was repaired by the hallux nail flap. Donor sites were repaired with artificial dermis in 6 patients and with ilioinguinal flaps in 10 patients. The effect of surgery was observed at outpatient clinic during postoperative follow-up. The survival of the hallux nail flap and the recovery of the donor site were observed. Function recovery were evaluated according to the Evaluation Standard of Finger Replantation and Reconstruction of Hand Surgery Society of Chinese Medical Association.Results:All the harvested hallux nail flaps matched with the profiles of recipient sites. All the hallux nail flaps survived over 4 to 30 months of follow-up, in an average of 16 months. Appearance of all hallux nail flaps was similar to normal thumbs, with good fingerprint and nails. Sensation recovery were S 3-S 4, with TPD at 4-7 mm. According to the Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association, 15 thumbs were excellent and 1 was good. Conclusion:Application of 3D printing assisted hallux nail flap transfer in reconstruction of defect of thumb in degloving injury can precisely design and harvest the required tissue and minimise a damage to the donor great toe. It improves the appearance of thumb as well as patient satisfaction. It is practical in reconstruction of the defect of thumb in degloving injury.
7.Accuracy and stability of a smart wearable device for quantitative analysis of environmental risks related to myopia
Ziyun WU ; Huanhuan HUO ; Yi LIU ; Xingrong WANG ; Yuanyuan HU ; Hongsheng BI
Chinese Journal of Experimental Ophthalmology 2023;41(8):746-754
Objective:To evaluate the accuracy and stability of Eye-Monitor, a smart wearable device, in quantifying environmental risks related to myopia.Methods:A diagnostic test study was conducted.Forty-two subjects aged 18-25 years old were recruited from Shandong University of Traditional Chinese Medicine in December 2021.Forty-two Eye-Monitors were selected from 80 devices using the simple random sampling method.Static and dynamic tests were carried out to compare environmental risks related to myopia.The static tests included measurements under different working distances, different head tilt angles when sitting, and different light intensities.The dynamic tests included measurements under different near-work time, different outdoor activity time, different time watching computers, and different phone viewing time.Eye-Monitor with the largest sum of absolute values of total relative error was selected, Spearman rank correlation analysis was used to analyze the correlation between the set values and Eye-Monitor measurements, and the accuracy of the objectively measured values was evaluated by Bland-Altman consistency analysis.The stability of the objectively measured values from Eye-Monitor was evaluated by the coefficient of variation.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of the Affiliated Eye Hospital of Shandong University of Traditional Chinese Medicine (No.HEC-KS-2021005KY). Written informed consent was obtained from each subject.Results:There were significant correlations between the measured values and setting values in the working distance, head tilt angle when sitting, indoor light intensity, total near-work time, average continuous near-work time, outdoor activity time, time watching computers, and time viewing phones ( rs=0.999, 0.998, 0.999, 0.998, 0.976, 0.959, 0.992, 0.997; all at P<0.001), with the 95% limits of agreement (LoA) of-1.23-2.32 cm, -1.49-4.24°, -13.90-26.90 lx, -6.46-0.11 minutes, -4.50-1.20 minutes, -4.01-1.34 minutes, -2.54-1.94 minutes and-2.15-0.45 minutes, respectively.More than 95% of dots were within the clinically acceptable LoA.The coefficients of variation of the measured values ranged from 1.23%-2.99%, 2.39%-8.25%, 0.87%-8.03%, 1.49%-12.52%, 6.63%-13.59%, 0.00%-14.15%, 1.20%-8.33 and 1.49%-12.51%, respectively, showing good stability.Eye-Monitor had good accuracy in measuring outdoor light intensity (95% LoA: -336.50-130.00 lx). Conclusions:The smart wearable device Eye-Monitor can be used to objectively monitor working distance, head tilt angle when sitting, indoor light intensity, total near-work time, average continuous near-work time, outdoor activity time, time watching computers, and time watching phones, which are with good accuracy and stability.
8.Identification of lipid droplets in gut bacteria.
Kai ZHANG ; Chang ZHOU ; Zemin LI ; Xuehan LI ; Ziyun ZHOU ; Linjia CHENG ; Ahmed Hammad MIRZA ; Yumeng SHI ; Bingbing CHEN ; Mengwei ZHANG ; Liujuan CUI ; Congyan ZHANG ; Taotao WEI ; Xuelin ZHANG ; Shuyan ZHANG ; Pingsheng LIU
Protein & Cell 2023;14(2):143-148
9.Clinical analysis of different parts of medullary infarction
Changyue LIU ; Yajing ZHANG ; Ziyun YUAN ; Chaonan LYU ; Peng DING ; Chenlu LI ; Huihui XUE ; Wei YUE
Chinese Journal of Neurology 2023;56(8):886-894
Objective:To investigate the clinical, imaging, etiological and prognostic features of patients with infarctions in different locations of the medulla oblongata.Methods:Patients with acute medullary infarction hospitalized at Tianjin Huanhu Hospital from July 2017 to July 2022 were included. The risk factors, clinical manifestation, stroke mechanism and 90-day prognosis of these patients were analyzed retrospectively.Results:Among the 256 patients enrolled, 150 (58.6%) had lateral medullary infarction (LMI), 106 (41.4%) had medial medullary infarction (MMI). The most frequent clinical manifestation of patients with LMI was dizziness (84.7%,127/150). And motor disorders (83.0%,88/106) was the most frequent clinical manifestation of patients with MMI. LMI lesions were mostly located in the middle (42.7%,64/150) and MMI lesions were mostly located in the upper (60.4%,64/106) medulla oblongata, with statistically significant difference (χ 2=47.53, P<0.001). Large artery atherosclerosis (LAA) was the main stroke mechanism in LMI and MMI [57.3%(86/150) vs 56.6%(60/106)]. Early neurological deterioration was more common in MMI (25.5%,27/106) and less common in LMI (7.3%,11/150), with statistically significant difference (χ 2=16.17, P<0.001). At discharge, more patients with MMI showed poor prognosis in short term [45.3% (48/106) vs 24.0% (36/150), with statistically significant difference (χ 2=12.76, P<0.001)] and even long term at 90-day follow-up [33.0% (35/106) vs 12.7% (19/150), also with statistically significant difference (χ 2=15.48, P<0.001)] than those with LMI. A total of 10 patients (4.0%, 10/256) developed respiratory failure during hospitalization, including 7 patients with LMI (4.7%, 7/150) and 3 patients with bilateral MMI (2.8%,3/106). Early neurological deterioration ( OR=3.38, 95% CI 1.25-9.10, P=0.016) and LAA (compared with small artery occlusion) ( OR=3.08, 95% CI 1.13-8.37, P=0.028) were independent risk factors for poor prognosis in MMI. Age ( OR=1.01, 95% CI 1.01-1.17, P=0.026) and early neurological deterioration ( OR=20.19, 95% CI=2.63-155.06, P=0.004) were independently correlated with poor outcome in LMI. Conclusions:LMI and MMI had similar etiology and significant differences in clinical manifestations, early neurological deterioration and prognosis. Further classification of medullary infarction was of great significance for diagnosis, treatment and prognosis evaluation.
10.Development and validation of an LC-MS/MS method for the determination of 12 ceramides in human plasma
Jinsong LIU ; Xiaoliang CHENG ; Ziyun HE ; Renqing YAN ; Wei ZHANG ; Shengkai YAN
Chinese Journal of Laboratory Medicine 2023;46(8):830-839
Objective:To establish and validate a reliable and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the detection of 12 ceramides in human plasma.Methods:From October 2021 to October 2022, 438 apparently healthy individuals were enrolled in the Affiliated Hospitals of Zunyi Medical University for reference intervals of 12 ceramides in this population. Plasma samples were collected, and separated using the ACQUITY UPLC BEH C18 (2.1×50 mm, 1.7 μm) column, deuterated isotopes were used as internal standards. The mobile phase is water (containing 0.1% formic acid) and isopropanol: acetonitrile (1∶1, v/v, containing 0.1% formic acid) at a flow rate of 0.4 ml/min with gradient elution. The detection method was established using the Qlife Lab 9000 Plus triple quadrupole mass spectrometer. The performance of the method was evaluated in terms of linearity, the lower limit of quantification, precision, recovery, and stability.Results:The method passed the performance evaluation in terms of linearity, the lower limit of quantification, recovery, precision, and stability. The intra-and inter-batch precision of the 12 ceramides ranged from 1.3% to 14.3%, the correctness was verified by spiked recovery experiments, and the recoveries ranged from 91.9% to 111.0%. The lower limit of quantification ranged from 0.001 to 0.100 μmol/L. Standard curve showed good linearity (correlation coefficient r>0.990). Stability tests showed that the 12 ceramides were stable in the biological matrix and after processing under different conditions for a specified period of time. The corresponding biological reference intervals were established for each of the 12 ceramides: 0.103-0.326 μmol/L for Cer(d18∶1/16∶0), 0.018-0.098 μmol/L for Cer(d18∶1/18∶0), 0.933-3.919 μmol/L for Cer(d18∶1/24∶0), 0.243-1.072 μmol/L for Cer(d18∶1/24∶1), 0.001-0.007 μmol/L for Cer(d18∶1/14∶0), 0.022-0.095 μmol/L for Cer(d18∶1/20∶0), 0.185-0.835 μmol/L for Cer(d18∶1/22∶0), 0.003-0.022 μmol/L for Cer(d18∶0/16∶0), 0.001-0.016 μmol/L for Cer(d18∶0/18∶0), 0.017-0.156 μmol/L for Cer(d18∶0/24∶0), 0.008-0.074 μmol/L for Cer(d18∶0/24∶1), and 0.106-0.721 μmol/L for LacCer(d18∶1/24∶1). Conclusion:Our study shows that the newly established LC-MS/MS method for the determination of 12 ceramides in human plasma is reliable, and suitable for clinical application.


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