1.A new strategy for pharmacodynamic substance screening and research on gut microbiota pathway mechanisms based on UPLC-Q-orbitrap-MS and 16S rRNA
Zhiying Yu ; Tong Li ; Jie Yang ; Jianghua He ; Weijiang Zhang ; Siyuan Li ; Yunpeng Qi ; Yihui Yin ; Ling Dong ; Wenjuan Xu
Journal of Traditional Chinese Medical Sciences 2025;2025(1):56-70
Objective:
To establish a progressive research strategy for “colonic components analysis - efficacy verification and mechanism exploration - gut microbiota”, screen pharmacodynamic substances, and investigate their mechanism via gut microbiota.
Methods:
The pharmacodynamics of Gegen Qinlian decoction (GQD) were assessed using a mouse model of dextran sulfate sodium-induced ulcerative colitis (UC). Ultra-performance liquid chromatography-quadrupole-orbitrap mass spectrometer was used to identify the prototype and metabolic components of GQD in the colon during UC. To analyze the structure and function of characteristic genera of GQD and its active components, 16S rRNA sequencing was performed.
Results:
We identified 67 prototypic and 14 metabolic components of GQD in the UC colon. The primary prototype components are flavonoids and alkaloids, including puerarin (PUE), baicalin (BAI), and berberine (BER). The metabolism was predominantly sulfonation. Efficacy verification showed that the main active components, puerarin, baicalin, and berberine, had good therapeutic effects on UC. The results of 16S rRNA gene sequencing showed that GQD improved UC by regulating the structure and function of the gut microbiota. The abundance of gut microbiota involved in the metabolism of the prototype components was influenced by the corresponding components. The function prediction results showed that PUE was the most comparable to GQD, with 24 consistent pathways. BAI and BER showed comparable gut microbiota regulation pathways. Characteristic pathways of BER include glucometabolic processes.
Conclusion
This study focused on the key issues in the gut microbiota pathway and developed a progressive research strategy to understand the transformation mechanisms of colonic components. This research systematically analyzed the active components and metabolic transformation of GQD in the colon during the pathological state of UC, as well as changes in the structure and function of the gut microbiota, clarified the mechanism of GQD and its active components in improving UC via the gut microbiota pathway.
2.Application of HPLC-MS/MS in the analysis of six tricarboxylic acid cycle metabolites in a mouse model of prenatal DEHP exposure
Wenkang Tao ; Lun Zhang ; Qianqian Huang ; Yun Yu ; Yue Jiang ; Mengzhen Hou ; Xutao Ling ; Fang Xie ; Jianqing Wang
Acta Universitatis Medicinalis Anhui 2025;60(5):897-905
Objective :
To establish a method for measuring major organic acids in the tricarboxylic acid cycle using a high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) system, and to investigate the changes in six tricarboxylic acid cycle organic acids(fumaric acid, malic acid, succinic acid, α-ketoglutaric acid, cis-aconitic acid, and citric acid) in the serum, liver, and placenta of mice exposed to di(2-ethylhexyl) phthalate(DEHP) during pregnancy.
Methods :
The serum, liver and placental samples from pregnant mice were processed and eluted through a Waters ACQUITY UPLC BEH Amide Column(130 Å, 1.7 μm, 2.1 mm × 150 mm) using a gradient elution program. Mobile phase A comprised an aqueous solution of 10 mmol/L ammonium acetate and 5 μmol/L methanephosphonic acid, while mobile phase B consisted of a 90% acetonitrile aqueous solution containing 10 mmol/L ammonium acetate and 5 μmol/L methanephosphonic acid, with a flow rate maintained at 0.35 ml/min. The mass spectrometry detection system utilized an electrospray ionization technique with negative ion mode for multiple reaction monitoring.
Results :
The correlation coefficients of the standard curves for the six tricarboxylic acid cycle organic acid metabolites were all above 0.996 within the quantitative range. The method's accuracy ranged from 97.14% to 108.26%, with inter-day and intra-day precision relative standard deviation between 1.35% and 6.73%. The matrix effect was between 93.29% and 107.47%, and the extraction recovery rate ranged from 94.82% to 112.57%. Analysis of six tricarboxylic acid cycle organic acids in the liver, serum, and placenta of DEHP-exposed mice during pregnancy showed significant reductions in fumaric acid, malic acid, α-ketoglutaric acid, cis-aconitic acid, and citric acid compared to the control group(P<0.05).
Conclusion
The HPLC-MS/MS method established in this study for detecting six tricarboxylic acid cycle organic acids in the serum, liver, and placenta of DEHP-exposed pregnant mice is stable, highly sensitive and selective. Prenatal DEHP exposure induced alterations in the levels of tricarboxylic acid(TCA) cycle organic acid metabolites in the liver, serum, and placenta of mice, suggesting that DEHP exposure during pregnancy may interfere with mitochondrial TCA cycle processes. These findings indicate potential value in the diagnosis and treatment of diseases associated with prenatal DEHP exposure.
3.Effect of stellate ganglion block on postoperative sleep quality in patients with surgery under general anesthesia:a meta analysis
Yu LIU ; Guangyou DUAN ; Ling DAN ; Yamei ZHANG
Chongqing Medicine 2025;54(1):181-186
Objective To evaluate the efficacy of stellate ganglion block(SGB)on postoperative sleep quality in the patients undergoing general anesthesia operation.Methods The randomized controlled trials(RCT)of the effect of SGB on postoperative sleep quality in the patients undergoing general anesthesia were searched from PubMed,Cochrane Library,Embase,CNKI,CBM,VIP and Wang Fang Data.The retrieval limi-tation was from the database establishment to July 21,2023.The screened literatures were evaluated by using the Cochrane 5.1.0 bias risk assessment tool,and then the meta analysis was performed by using Rev Man 5.4 software.Results A total of eight literatures were screened and included.The results of meta analysis showed that the incidence rate of sleep disorders in the first night after surgery in the SGB group was lower than that in the control group(OR=0.26,95%CI:0.15-0.45,P<0.001),and the postoperative total sleep duration in the SGB group was higher than that in the control group(MD=69.06,95%CI:53.69-84.44,P<0.001),the sleep efficiency of the SGB group was higher than that of the control group(MD=13.59,95%CI:10.70-16.49,P<0.001);the postoperative Assens Insomnia Scale(AIS)scores(MD=-1.73,95%CI:-2.43 to-1.03,P<0.001)and Pittsburgh Sleep Quality Index Scale(PSQI)scores(MD=-3.26,95%CI:-5.40 to-1.12,P<0.003)in the SGB group were lower than those in the control group.Conclusion SGB could reduce the incidence rate of sleep disturbances at the postoperative first night,extend the postoperative total sleep time,increase the postoperative sleep efficiency,reduce the postoperative sleep quality score,and improve the postoperative sleep quality,which is a non-pharmacological method with good effect and high safety,has good clinical application prospect and is worth promoting.
4.Study on the predictive value of ICH-LR2S2 score and ICH-APS score for stroke-associated pneumonia in patients with spontaneous intracerebral hemorrhage
Yan LIU ; Xiumei ZOU ; Lishang LIAO ; Jianfei HUANG ; Ling ZHANG ; Yu RAN ; Guangzhen LI
Chongqing Medicine 2025;54(5):1111-1117
Objective To study the predictive value of the Prognostic Assessment Scale for Patients with Spontaneous Intracerebral Hemorrhage(ICH-LR2S2)and the Prehospital Risk Assessment Scale for Prehospital Deterioration Risk Assessment Scale(ICH-APS)for the development of stroke-associated pneu-monia(SAP)in patients with spontaneous intracerebral hemorrhage.Methods A total of 349 patients with spontaneous intracerebral hemorrhage who were hospitalized for the first time in this hospital from July 2023 to July 2024 were selected as the research subjects.The general demographic data and medical documentations of the patients were collected,and ICH-LR2S2 score and ICH-APS score were carried out within 48 hours after admission.According to whether pneumonia occurred within 7 days after admission,the patients were divided into the SAP group and the non-SAP group,and the diagnostic efficiency of the ICH-LR2S2 score and ICH-APS score for SAP in patients with spontaneous intracerebral hemorrhage was evaluated.Results Among the 349 patients with spontaneous intracerebral hemorrhage,98 patients(28.08%)had pneumonia.The results of multivariate logistic regression analysis showed that age,history of chronic obstructive pulmonary disease(COPD),nasogastric tube,tracheal intubation,National Institutes of Health Stroke Scale(NIHSS)score,Glasgow Coma Scale(GCS)score,C-reactive protein,fasting blood glucose,dysphagia,ICH-LR2S2 score,ICH-APS-A score,and ICH-APS-B score were independent influencing factors for SAP in patients with spontane-ous intracerebral hemorrhage(P<0.05).The results of the receiver operating characteristic(ROC)curve showed that the ICH-LR2S2 score had the highest diagnostic efficiency for SAP in patients with spontaneous intracerebral hemorrhage,with an area under the curve(AUC)of 0.837,a sensitivity of 0.827,a specificity of 0.783,and a Youden index of 0.610.Conclusion ICH-LR2S2 score has a high predictive value for the occur-rence of SAP in patients with spontaneous intracerebral hemorrhage.
5.Armored IL15 or IL15 receptor adaptors enhance proliferation and anti-tumor activity of CD19-targeted CAR-NK cells
Mengyuan YU ; Xiuying LIU ; Xiaotian ZHANG ; Jiaying WANG ; Zhiming LING ; Jianxun WANG
Journal of Army Medical University 2025;47(23):2903-2912
Objective To construct CD19-targeted chimeric antigen receptor natural killer(CAR-NK)cells armored with interleukin15(IL15)or IL15 receptor-linker(RLI)and preliminarily validate their proliferative capacity and anti-tumor activity in vitro.Methods Natural killer cells(NK92 and NK92MI)from patients with human malignant non-Hodgkin lymphoma were cultured,and CD19-targeted CAR-NK cells armored with IL15 or RLI were prepared using retroviral vector particles.IL15 secretion was measured by ELISA,and proliferative capacity was assessed via CFSE dilution assays.Human B-lymphocytic leukemia cells(Nalm6-GFP-Luc)and human colon cancer cells overexpressing CD19(hCD19-SW620-GFP-Luc)were cultured,with surface CD19 expression confirmed(>99%positivity for both).Anti-tumor activity was evaluated by measuring cytotoxicity at effector-to-target(E:T)ratios using luciferase-based assays(4/12 h),detecting surface CD107a expression via flow cytometry,and quantifying cytokine release using CBA assays.Results NK92/NK92MI-CD19 CAR cells armored with IL15 or RLI were successfully generated.IL15 secretion was significantly higher in armored groups versus non-armored controls(P<0.01).Without IL-2 stimulation,IL15/RLI enhanced CAR-NK proliferation(P<0.05).Both armored designs significantly increased tumor-killing efficiency(P<0.05)and CD107a degranulation.IL15/RLI-armored NK92-CD19 CAR cells exhibited elevated release of IL2,IL10,IL6,TNF-α,sFas,IFN-γ,and Granulysin(P<0.05),while armored NK92MI-CD19 CAR cells showed additional increases in Granzyme A,Granzyme B,and Perforin(P<0.05).Conclusion IL15/RLI-armored NK92/NK92MI-CD19 CAR cells demonstrate potent anti-tumor activity,supporting their combined clinical therapeutic potential for tumors.
6.Atomic Fluorescence Dispersion Detection Technique Based on Area Array Single Photon Counting Imaging Detector
Chen TAO ; Hong-Ji ZHANG ; Chun-Sheng LI ; Ling-Ping HE ; Zhen-Yu MA ; Bo CHEN ; Ran ZHANG
Chinese Journal of Analytical Chemistry 2025;53(2):187-194
The single photon counting imaging detector based on microchannel plate(MCP)has the characteristics of high sensitivity and low dark count rate,and has been applied to the optical remote sensing detection of weak ultraviolet spectral signals in space.In this work,by using planar array single photon counting imaging detector as the detector,flat-field concave grating as the splitter,and electrodeless discharge lamp(EDL)as the excitation light source,a dispersion detection system suitable for hydride generation-atomic fluorescence spectrometry(HG-AFS)was developed.The wavelength calibration of the system was carried out,and the negative high pressure and EDL stability time of the planar array single photon counting imaging detector were analyzed and optimized.The characteristic emission spectral lines of As and Bi elements excited in the wavelength range of 180-320 nm were analyzed,and the scattering interference in the wavelength range of 257.3-306.7 nm was discussed.The results showed that the AFS dispersion detection system based on the planar array single photon counting imaging detector could detect and analyze the HG-AFS fluorescence signal initially,and the influence of scattering interference on the detection results was effectively avoided.The system had the advantages including simple structure,no refrigeration and temperature control,no moving parts and simultaneous measurement of multi-band.
7.Ultrasonic Extraction-Single-Particle Inductively Coupled Plasma Mass Spectrometry for Analysis of Metal Nanoparticles in Seawater Sediments
Jie CHUN ; Yu-Mei SONG ; Chang WANG ; Rui-Ling ZHANG ; Peng-Ran GUO ; Wei-Xin LIANG ; Ting ZHANG
Chinese Journal of Analytical Chemistry 2025;53(6):987-997
Accurate analysis of metal nanoparticles(MNPs)in sediments is a prerequisite for assessing the ecological risks of MNPs in aquatic environmental sediments.In this study,an analytical method for quantitative detection of concentration and particle size distribution of silver-containing nanoparticles(Ag-NPs),zinc-containing nanoparticles(Zn-NPs),cerium-containing nanoparticles(Ce-NPs),and titanium-containing nanoparticles(Ti-NPs)in sediments was established based on ultrasonic extraction-single particle inductively coupled plasma mass spectrometry(SP-ICP-MS).The effects of sample preparation conditions such as extraction solvent type,solid-liquid ratio,ultrasonic time,and settling time on the recovery of MNPs were investigated.The results showed that the extraction of MNPs from sediment by distilled water could effectively eliminate the high background signal interference introduced by the extractant under the conditions of solid-liquid ratio of 1∶400(g∶mL),ultrasonic extraction time of 1 h and settling time of 3 h.The detection limits for particle size of Ag-NPs,Zn-NPs,Ce-NPs and Ti-NPs in sediments were 31,35,26 and 85 nm,respectively,while the detection limits of particle concentrations were 1.21×104,1.90×104,5.26×107 and 1.48×107 particles/g,respectively.The spiking recoveries of Ag-NPs,Zn-NPs,Ce-NPs and Ti-NPs in sediments were 62.1%-108.7%,with relative standard deviations below 10%.This method could rapidly,accurately and simultaneously determine the concentration and particle size distribution of various MNPs in sediments,and was successfully applied to analysis of Ag-NPs,Zn-NPs,Ce-NPs,and Ti-NPs in authentic marine sediments.
8.Progress of Microfluidics-Integrated Clustered Regularly Interspaced Short Palindromic Repeats Detection Systems for Pathogen Diagnosis in Point-of-Care Testing
Qian XU ; Jing ZHANG ; Yu-Ling QIN ; Li WU
Chinese Journal of Analytical Chemistry 2025;53(11):1761-1772
Clustered regularly interspaced short palindromic repeats-associated proteins(CRISPR-Cas)system has emerged as a next-generation nucleic acid detection tool due to its precise gene editing capabilities and exhibits high specificity and significant potential for application in the diagnosis of pathogen.However,the inherent shortcomings of CRISPR-Cas,including insufficient sensitivity of the traditional CRISPR-Cas system,complex operation of the CRISPR-Cas system combined with amplification,and limited capability for multi-target detection,have significantly constrained its practical applications in clinical settings.With the advantages of miniaturization,integration and automation,microfluidics has emerged as a promising tool for advancing the development of point-of-care testing(POCT)platforms.The integration of microfluidic technology with CRISPR-Cas system not only enhances the detection throughput and efficiency significantly,but also drives innovation in the development of portable POCT devices.In this paper,a comprehensive review of the research advancements of CRISPR-Cas nucleic acid detection platform facilitated by microfluidic technology was presented,with a focus on their application in detection of pathogens associated with clinical disease.This study aimed to explore emerging molecular diagnostic methodologies and offered novel insights for future research in clinical pathogen detection.
9.Improvement effect of rehabilitation nursing based on IKAP theory on patients with urinary incontinence after radical prostatectomy.
Ting-Ting XIA ; Wen-Fang CHEN ; Jie LIU ; Xiao-Wen TAN ; Juan LI ; Yan-Yan ZHANG ; Yu-Mei CAO ; Song XU ; Ting-Ling ZHANG
National Journal of Andrology 2025;31(5):438-443
OBJECTIVE:
To explore the improvement effect of rehabilitation nursing based on information-knowledge-belief-behavior (IKAP) theory on urinary incontinence patients after radical prostatectomy.
METHODS
Sixty-six patients with urinary incontinence who received robot-assisted laparoscopic radical prostatectomy in General Hospital of Eastern Theater Command from January 2021 to January 2023 were selected and divided into control group (n=33) and observation group (n=33) according to random number table method. The patients in the control group were treated with rehabilitation nursing. The patients in the observation group were treated with rehabilitation nursing guided by IKAP theory. The recovery of urinary incontinence, duration of urinary incontinence, subjective well-being, quality of life, psychological and emotional indexes of patients in the two groups were compared. Results: The total effective rate of urinary incontinence recovery in the observation group was significantly higher than that in the control group (90.91% vs 60.61%,P<0.05). The duration of urinary incontinence in the observation group was significantly shorter than that in the control group ([3.36±1.54]d vs [4.15±1.36]d,P<0.05). And the subjective well-being score in observation group was significantly higher than that in the control group ([19.36±2.69]points vs [11.65±2.65]points, P<0.05). There was no significant difference in preoperative physical function, social function,and mental health scores between the two groups (P>0.05). And all scores in the observation group were significantly higher than those in the control group after surgery (P<0.05). There was no significant difference in the preoperative SAS and SDS scores between the two groups of patients (P>0.05). And the scores of SAS and SDS in observation group were lower than those of the control group after the operation (P<0.05). Conclusion: Rehabilitation nursing based on IKAP theory can significantly improve urinary incontinence in patients with prostate cancer after surgery, which promotes the recovery of urinary incontinence, shortens the time of urinary incontinence, and improves the subjective well-being and quality of life, as well as reduces the negative impact of negative emotions. Therefore, it can be widely promoted and implemented in clinical practice.
Humans
;
Prostatectomy/adverse effects*
;
Urinary Incontinence/etiology*
;
Male
;
Quality of Life
;
Rehabilitation Nursing
;
Middle Aged
;
Aged
10.Association of short-term air pollution with risk of major adverse cardiovascular event mortality and modification effects of lifestyle in Chinese adults.
Wendi XIAO ; Xin YAO ; Yinqi DING ; Junpei TAO ; Canqing YU ; Dianjianyi SUN ; Pei PEI ; Ling YANG ; Yiping CHEN ; Huaidong DU ; Dan SCHMIDT ; Yaoming ZHAI ; Junshi CHEN ; Zhengming CHEN ; Jun LV ; Liqiang ZHANG ; Tao HUANG ; Liming LI
Environmental Health and Preventive Medicine 2025;30():38-38
BACKGROUND:
Previous evidence showed that ambient air pollution and cardiovascular mortality are related. However, there is a lack of evidence towards the modification effect of long-term lifestyle on the association between short-term ambient air pollution and death from cardiovascular events.
METHOD:
A total of 14,609 death from major adverse cardiovascular events (MACE) were identified among the China Kadoorie Biobank participants from 2013 to 2018. Ambient air pollution exposure including particulate matter 2.5 (PM2.5), SO2, NO2, CO, and O3 from the same period were obtained from space-time model reconstructions based on remote sensing data. Case-crossover design and conditional logistic regression was applied to estimate the effect of short-term exposure to air pollutants on MACE mortality.
RESULTS:
We found MACE mortality was significantly associated with PM2.5 (relative percent increase 2.91% per 10 µg/m3 increase, 95% CI 1.32-4.53), NO2 (5.37% per 10 µg/m3 increase, 95% CI 1.56-9.33), SO2 (6.82% per 10 µg/m3 increase, 95% CI 2.99-10.80), and CO (2.24% per 0.1 mg/m3 increase, 95% CI 1.02-3.48). Stratified analyses indicated that drinking was associated with elevated risk of MACE mortality with NO2 and SO2 exposure; physical inactivity was associated with higher risk of death from MACE when exposed to PM2.5; and people who had balanced diet had lower risk of MACE mortality when exposed to CO and NO2.
CONCLUSIONS
The study results showed that short-term exposure to ambient PM2.5, NO2, SO2, and CO would aggravate the risk of cardiovascular mortality, yet healthy lifestyle conduct might mitigate such negative impact to some extent.
Humans
;
Cardiovascular Diseases/epidemiology*
;
China/epidemiology*
;
Male
;
Female
;
Air Pollution/adverse effects*
;
Middle Aged
;
Air Pollutants/analysis*
;
Particulate Matter/analysis*
;
Environmental Exposure/adverse effects*
;
Life Style
;
Aged
;
Adult
;
Risk Factors
;
Cross-Over Studies
;
East Asian People


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