1.Quality evaluation of Qingwen hufei granules based on fingerprints combined with multi-component content determination
Huiying ZHOU ; Yuan WANG ; Yani WANG ; Yun YANG ; Bo WANG ; Shuanzhu YANG ; Liping CAO ; Hong ZHANG ; Kaihua LONG
China Pharmacy 2026;37(3):338-343
OBJECTIVE To provide a scientific basis for the quality evaluation and clinical application of Qingwen hufei granules. METHODS Fourteen batches of Qingwen hufei granules were used as samples to establish high-performance liquid chromatography (HPLC) fingerprints using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (2012 Edition). The chromatographic peaks were identified and the similarity was evaluated. Cluster analysis (CA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used to conduct chemical pattern recognition analysis on the 14 batches of samples. Meanwhile, the contents of neochlorogenic acid (NGA), chlorogenic acid (CHA), cryptochlorogenic acid (CGA), forsythoside A (FTA), 3,5-O-dicaffeoylquinic acid (3,5-O- DA), 4,5-O-dicaffeoylquinic acid (4,5-O-DA), and angoroside C (AGC) in the samples were determined by HPLC. RESULTS The methodological investigation results of both the fingerprint and the content determination complied with the relevant requirements. Fourteen common peaks were indicated in the HPLC fingerprints of the 14 batches of samples, and 7 of them were identified [NGA (peak 2), CHA (peak 3), CGA (peak 5), FTA (peak 11), 3,5-O-DA (peak 12), 4,5-O-DA (peak 13), and AGC (peak 14)]; the similarity of each sample was greater than 0.94. The results of CA and PCA showed that the samples could be classified into 3 categories; the results of OPLS-DA indicated that peak 4 (unknown), peak 11 (FTA), peak 8 (unknown), peak 9 (unknown), and peak 1 (unknown) were the differential components. The content ranges of NGA, CHA, CGA, 3,5-O-DA, FTA, 4,5-O-DA and AGC in the 14 batches of samples were 0.210 4-0.458 7, 0.269 1-0.506 3, 0.228 1-0.461 1, 0.443 9-1.044 6, 0.066 7-0.155 7, 0.062 8-0.143 8, and 0.057 4-0.105 7 mg/g, respectively. CONCLUSIONS The HPLC fingerprint and multi-component content determination methods established in this study are efficient and reliable, and can be used for the quality evaluation of Qingwen hufei granules.
2.Exploring on Quality Evaluation Methods of Clinical Case Reports in Traditional Chinese Medicine Based on China Clinical Cases Library of Traditional Chinese Medicine
Kaige ZHANG ; Feng ZHANG ; Bo ZHOU ; Haimin CHEN ; Yong ZHU ; Changcheng HOU ; Liangzhen YOU ; Weijun HUANG ; Jie YANG ; Guoshuang ZHU ; Shukun GONG ; Jianwen HE ; Yang YE ; Yuqiu AN ; Chunquan SUN ; Qingjie YUAN ; Buman LI ; Xingzhong FENG ; Kegang CAO ; Hongcai SHANG ; Jihua GUO ; Xiaoxiao ZHANG ; Zhining TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):271-276
As the core vehicle for preserving and transmitting traditional Chinese medicine(TCM) academic thought and clinical experience, the establishment of a robust quality evaluation system for TCM clinical case reports is a crucial component in the current standardization and modernization of TCM. Based on the practical experience of constructing the China Clinical Cases Library of Traditional Chinese Medicine by the China Association of Chinese Medicine, this study conducted a comprehensive analysis of critical challenges, including insufficient authenticity and unfocused evaluation criteria. It proposed a three-dimensional evaluation framework grounded in the structure-process-outcome logic, encompassing three dimensions of authenticity and standardization, characteristics and advantages, application and translational impact. This framework integrated 12 key evaluation indicators in a systematic manner. The model preserved the academic characteristics of TCM syndrome differentiation and treatment, while aligning with modern scientific research standards, achieving a balance between individualized TCM experience and standardized evaluation. Concurrently, this study provided theoretical foundations and methodological guidance for evaluating the quality of TCM clinical cases, contributing significantly to the inheritance of TCM knowledge, evidence-based practice, and the reform of talent evaluation mechanisms.
3.Mechanisms of Salvianolic Acid B in Inhibiting Epithelial-mesenchymal Transition in Non-small Cell Lung Cancer by Downregulating PAICS Expression
Bo XU ; Jixian ZHANG ; Linling HU ; Bo JIANG ; Shasha YUAN ; Yiling FAN ; Zhishen RUAN ; Yihan YU ; Qing MIAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):23-33
ObjectiveTo investigate the molecular mechanisms by which salvianolic acid B (SalB) inhibits epithelial-mesenchymal transition (EMT) in non-small cell lung cancer (NSCLC) by downregulating phosphoribosylaminoimidazole carboxylase and phosphoribosylaminoimidazole succinocarboxamide synthetase (PAICS) expression. MethodsNSCLC A549 cells and normal bronchial epithelial cells (bronchial epithelium transformed with Ad12-SV40 2B, BEAS-2B) were used as models. Cell viability was assessed using the cell counting kit-8 (CCK-8) assay after treatment with SalB (0, 50, 100, 200, 300, 400, 500 μmol·L-1 for 24 or 48 h to determine effective and safe intervention concentrations. Cell proliferation, cell cycle distribution, and apoptosis were evaluated by 5-ethynyl-2′-deoxyuridine (EdU) staining and flow cytometry, respectively. Wound healing and Transwell invasion assays were performed to assess cell migration and invasion. RNA sequencing combined with bioinformatic analysis was conducted to identify differentially expressed genes and functional enrichment. Molecular docking was used to predict the binding ability between SalB and PAICS, and the cellular thermal shift assay (CETSA) was performed to evaluate the effect of SalB on the thermal stability of the PAICS protein. Western blot (WB) was used to detect the effects of SalB on PAICS and EMT-related proteins (E-cadherin, N-cadherin, Vimentin, Snail, and Slug). A functional rescue assay was conducted by PAICS overexpression via plasmid transfection. ResultsCompared with the control group, SalB inhibited A549 cell viability in a dose-dependent manner (P<0.05), and the effective concentrations (≤300 μmol·L-1) showed no significant cytotoxicity in BEAS-2B cells. Within this concentration range, SalB significantly inhibited A549 cell proliferation, migration, and invasion, and induced G0/G1 phase arrest and apoptosis (P<0.05). Transcriptomic analysis showed that SalB significantly downregulated PAICS expression, and its functions were enriched in cell proliferation and EMT. Bioinformatic analysis indicated that PAICS is highly expressed in lung adenocarcinoma and is associated with poor prognosis (P<0.01). Molecular docking showed that SalB has strong binding ability to PAICS (binding energy -9.1 kcal·mol-1. CETSA results showed that SalB significantly increased the thermal stability of the PAICS protein (P<0.05). WB results showed that, compared with the control group, SalB dose-dependently downregulated PAICS expression, upregulated E-cadherin, and downregulated N-cadherin, Vimentin, Snail, and Slug (P<0.05). Functional rescue experiments showed that, compared with the empty vector group, PAICS overexpression significantly enhanced A549 cell proliferation, migration, and invasion, promoted cell cycle progression, and inhibited apoptosis (P<0.05). Meanwhile, compared with the empty vector + SalB-H group, PAICS overexpression partially reversed the inhibitory effects of SalB on malignant phenotypes and EMT-related proteins (N-cadherin, Vimentin, Snail, and Slug), and downregulated E-cadherin expression (P<0.05,P<0.01), indicating that PAICS is a key functional target mediating the antitumor effects of SalB. ConclusionSalB effectively inhibits EMT progression and cell cycle progression in A549 cells by downregulating PAICS expression, thereby exerting anti-NSCLC effects. This study not only reveals that PAICS is a key functional target through which SalB regulates EMT, but also provides experimental evidence supporting SalB as a potential candidate drug for inhibiting NSCLC metastasis.
4.Eupatilin Inhibits Proliferation, Invasion, and Metastasis of Non-small Cell Lung Cancer via EZH2/H3K27me3 Signaling Pathway
Bo XU ; Yihan YU ; Linling HU ; Bo JIANG ; Yu QI ; Shasha YUAN ; Yiling FAN ; Jixian ZHANG ; Qing MIAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):58-69
ObjectiveTo investigate the mechanisms by which eupatilin (Eup) inhibits proliferation, invasion, and metastasis of non-small cell lung cancer (NSCLC) through the enhancer of zeste homolog 2/histone H3 lysine 27 trimethylation (EZH2/H3K27me3) signaling pathway. MethodsIn vivo, a subcutaneous xenograft tumor model was established in nude mice using H1299 cells to evaluate the anti-NSCLC effects of Eup. Immunohistochemistry (IHC-P) was used to detect the expression of proliferation- and invasion/metastasis-related proteins, including proliferating cell nuclear antigen (PCNA), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), and vascular endothelial growth factor A (VEGFA). In vitro, cell counting kit-8 (CCK-8) assays were performed to determine the viability of H1299 cells treated with different concentrations of Eup (0-200 μmol·L-1) and to select appropriate concentrations. Colony formation and 5-ethynyl-2′-deoxyuridine (EdU) assays were used to evaluate cell proliferation. Wound healing and invasion assays were conducted to assess cell migration and invasion. Human umbilical vein endothelial cell (HUVEC) angiogenesis assays were used to evaluate the effects of Eup on angiogenesis. Transcriptomic analysis was performed to identify the targets of Eup in H1299 cells and to explore its major functions. Molecular docking and molecular dynamics simulations were conducted to predict the binding affinity and interaction stability between Eup and its target proteins. Western blot was used to detect the effects of Eup on the expression levels of EZH2/H3K27me3 pathway-related proteins and proliferation- and invasion/metastasis-related proteins, including PCNA, MMP-2, MMP-9, and VEGFA. ResultsIn the subcutaneous xenograft model, compared with the model group, Eup treatment dose-dependently inhibited the growth of H1299 xenograft tumors, and the tumor inhibition rate was significantly increased (P<0.05). IHC-P results showed that, compared with the model group, high-dose Eup significantly reduced the expression levels of PCNA, MMP-2, MMP-9, and VEGFA in vivo (P<0.05). In vitro, compared with the control group, Eup inhibited the proliferation, invasion, and metastasis of NSCLC cells in a concentration-dependent manner. Transcriptomic analysis further showed that, compared with the control group, Eup significantly downregulated EZH2 expression, and its functional effects were associated with inhibition of tumor metastasis. Molecular docking and molecular dynamics simulations indicated that Eup exhibited strong binding affinity with EZH2 and stable interactions. Western blot results demonstrated that, compared with the model group, Eup significantly inhibited, in a dose-dependent manner, the expression levels of EZH2, H3K27me3, and proliferation- and invasion/metastasis-related proteins (PCNA, MMP-2, MMP-9, and VEGFA) in both in vivo and in vitro experiments (P<0.05). In vitro, compared with the control group, overexpression of EZH2 via plasmid transfection partially reversed the inhibitory effects of Eup on the expression of key proteins involved in proliferation and invasion/metastasis in H1299 cells. ConclusionEup effectively inhibits the proliferation, migration, and invasion of H1299 cells both in vivo and in vitro. The underlying mechanism may be related to inhibition of the EZH2/H3K27me3 signaling pathway and downregulation of proliferation- and invasion/metastasis-related proteins, including PCNA, MMP-2, MMP-9, and VEGFA. Eup may serve as a potential therapeutic agent for suppressing proliferation and invasion/metastasis in NSCLC.
5.Mechanisms of Salvianolic Acid B in Inhibiting Epithelial-mesenchymal Transition in Non-small Cell Lung Cancer by Downregulating PAICS Expression
Bo XU ; Jixian ZHANG ; Linling HU ; Bo JIANG ; Shasha YUAN ; Yiling FAN ; Zhishen RUAN ; Yihan YU ; Qing MIAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):23-33
ObjectiveTo investigate the molecular mechanisms by which salvianolic acid B (SalB) inhibits epithelial-mesenchymal transition (EMT) in non-small cell lung cancer (NSCLC) by downregulating phosphoribosylaminoimidazole carboxylase and phosphoribosylaminoimidazole succinocarboxamide synthetase (PAICS) expression. MethodsNSCLC A549 cells and normal bronchial epithelial cells (bronchial epithelium transformed with Ad12-SV40 2B, BEAS-2B) were used as models. Cell viability was assessed using the cell counting kit-8 (CCK-8) assay after treatment with SalB (0, 50, 100, 200, 300, 400, 500 μmol·L-1 for 24 or 48 h to determine effective and safe intervention concentrations. Cell proliferation, cell cycle distribution, and apoptosis were evaluated by 5-ethynyl-2′-deoxyuridine (EdU) staining and flow cytometry, respectively. Wound healing and Transwell invasion assays were performed to assess cell migration and invasion. RNA sequencing combined with bioinformatic analysis was conducted to identify differentially expressed genes and functional enrichment. Molecular docking was used to predict the binding ability between SalB and PAICS, and the cellular thermal shift assay (CETSA) was performed to evaluate the effect of SalB on the thermal stability of the PAICS protein. Western blot (WB) was used to detect the effects of SalB on PAICS and EMT-related proteins (E-cadherin, N-cadherin, Vimentin, Snail, and Slug). A functional rescue assay was conducted by PAICS overexpression via plasmid transfection. ResultsCompared with the control group, SalB inhibited A549 cell viability in a dose-dependent manner (P<0.05), and the effective concentrations (≤300 μmol·L-1) showed no significant cytotoxicity in BEAS-2B cells. Within this concentration range, SalB significantly inhibited A549 cell proliferation, migration, and invasion, and induced G0/G1 phase arrest and apoptosis (P<0.05). Transcriptomic analysis showed that SalB significantly downregulated PAICS expression, and its functions were enriched in cell proliferation and EMT. Bioinformatic analysis indicated that PAICS is highly expressed in lung adenocarcinoma and is associated with poor prognosis (P<0.01). Molecular docking showed that SalB has strong binding ability to PAICS (binding energy -9.1 kcal·mol-1. CETSA results showed that SalB significantly increased the thermal stability of the PAICS protein (P<0.05). WB results showed that, compared with the control group, SalB dose-dependently downregulated PAICS expression, upregulated E-cadherin, and downregulated N-cadherin, Vimentin, Snail, and Slug (P<0.05). Functional rescue experiments showed that, compared with the empty vector group, PAICS overexpression significantly enhanced A549 cell proliferation, migration, and invasion, promoted cell cycle progression, and inhibited apoptosis (P<0.05). Meanwhile, compared with the empty vector + SalB-H group, PAICS overexpression partially reversed the inhibitory effects of SalB on malignant phenotypes and EMT-related proteins (N-cadherin, Vimentin, Snail, and Slug), and downregulated E-cadherin expression (P<0.05,P<0.01), indicating that PAICS is a key functional target mediating the antitumor effects of SalB. ConclusionSalB effectively inhibits EMT progression and cell cycle progression in A549 cells by downregulating PAICS expression, thereby exerting anti-NSCLC effects. This study not only reveals that PAICS is a key functional target through which SalB regulates EMT, but also provides experimental evidence supporting SalB as a potential candidate drug for inhibiting NSCLC metastasis.
6.Eupatilin Inhibits Proliferation, Invasion, and Metastasis of Non-small Cell Lung Cancer via EZH2/H3K27me3 Signaling Pathway
Bo XU ; Yihan YU ; Linling HU ; Bo JIANG ; Yu QI ; Shasha YUAN ; Yiling FAN ; Jixian ZHANG ; Qing MIAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):58-69
ObjectiveTo investigate the mechanisms by which eupatilin (Eup) inhibits proliferation, invasion, and metastasis of non-small cell lung cancer (NSCLC) through the enhancer of zeste homolog 2/histone H3 lysine 27 trimethylation (EZH2/H3K27me3) signaling pathway. MethodsIn vivo, a subcutaneous xenograft tumor model was established in nude mice using H1299 cells to evaluate the anti-NSCLC effects of Eup. Immunohistochemistry (IHC-P) was used to detect the expression of proliferation- and invasion/metastasis-related proteins, including proliferating cell nuclear antigen (PCNA), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), and vascular endothelial growth factor A (VEGFA). In vitro, cell counting kit-8 (CCK-8) assays were performed to determine the viability of H1299 cells treated with different concentrations of Eup (0-200 μmol·L-1) and to select appropriate concentrations. Colony formation and 5-ethynyl-2′-deoxyuridine (EdU) assays were used to evaluate cell proliferation. Wound healing and invasion assays were conducted to assess cell migration and invasion. Human umbilical vein endothelial cell (HUVEC) angiogenesis assays were used to evaluate the effects of Eup on angiogenesis. Transcriptomic analysis was performed to identify the targets of Eup in H1299 cells and to explore its major functions. Molecular docking and molecular dynamics simulations were conducted to predict the binding affinity and interaction stability between Eup and its target proteins. Western blot was used to detect the effects of Eup on the expression levels of EZH2/H3K27me3 pathway-related proteins and proliferation- and invasion/metastasis-related proteins, including PCNA, MMP-2, MMP-9, and VEGFA. ResultsIn the subcutaneous xenograft model, compared with the model group, Eup treatment dose-dependently inhibited the growth of H1299 xenograft tumors, and the tumor inhibition rate was significantly increased (P<0.05). IHC-P results showed that, compared with the model group, high-dose Eup significantly reduced the expression levels of PCNA, MMP-2, MMP-9, and VEGFA in vivo (P<0.05). In vitro, compared with the control group, Eup inhibited the proliferation, invasion, and metastasis of NSCLC cells in a concentration-dependent manner. Transcriptomic analysis further showed that, compared with the control group, Eup significantly downregulated EZH2 expression, and its functional effects were associated with inhibition of tumor metastasis. Molecular docking and molecular dynamics simulations indicated that Eup exhibited strong binding affinity with EZH2 and stable interactions. Western blot results demonstrated that, compared with the model group, Eup significantly inhibited, in a dose-dependent manner, the expression levels of EZH2, H3K27me3, and proliferation- and invasion/metastasis-related proteins (PCNA, MMP-2, MMP-9, and VEGFA) in both in vivo and in vitro experiments (P<0.05). In vitro, compared with the control group, overexpression of EZH2 via plasmid transfection partially reversed the inhibitory effects of Eup on the expression of key proteins involved in proliferation and invasion/metastasis in H1299 cells. ConclusionEup effectively inhibits the proliferation, migration, and invasion of H1299 cells both in vivo and in vitro. The underlying mechanism may be related to inhibition of the EZH2/H3K27me3 signaling pathway and downregulation of proliferation- and invasion/metastasis-related proteins, including PCNA, MMP-2, MMP-9, and VEGFA. Eup may serve as a potential therapeutic agent for suppressing proliferation and invasion/metastasis in NSCLC.
7.Mosquito monitoring and influence of meteorological factors on mosquito density around Beijing Capital International Airport, 2016-2022
Ran FENG ; Tie-zheng MA ; Si-jie ZHU ; Bo TIAN ; Fei QUAN ; Zhi-lin WU ; Xiao-tao LIU ; Fu-yuan ZHANG ; Song-jian ZHANG
Acta Parasitologica et Medica Entomologica Sinica 2026;33(1):40-48
Objective To understand mosquito density, species composition, and seasonal variations around the Beijing Capital International Airport(BCIA)and provide scientific evidence for mosquito-borne disease prevention and control. Methods Meteorological data were collected from 2016 to 2022, and mosquito density was monitored at seven surveillance sites around the BCIA from May to October each year using CO2-baited mosquito traps to analyze the relationship between mosquito density and meteorological factors. Results In total,68 518 female mosquitoes were captured, with a mosquito density of 10.20 per light · hour. The dominant species was Culex pipiens pallens(73.90%), followed by Aedes albopictus(9.71%)and Ae. vexans(9.29%);the mosquito density was highest in 2016(14.8 per light·hour)and lowest in 2022(4.46 per light·hour). Significant statistical differences were observed in mosquito density among different species(F=18.118, P<0.05). The density of Cx. pipiens pallens and other mosquito species showed a mutually exclusive trend; the peak of mosquito density varied based on years and habitats, with significant statistical differences in mosquito density among different habitats(F=8.504, P<0.05). The highest mosquito density was observed near the Wenyu River(21.50 per light·hour), whereas the lowest was recorded at the BCIA construction site(2.44 per light· hour). The monthly average temperature(r=0.595), monthly average highest temperature(r=0.575), and monthly average lowest temperature(r=0.624)showed moderate positive correlations, whereas the monthly average air pressure(r=-0.484)showed a moderate negative correlation. The monthly average minimum temperature was included in the regression model(F=25.575, P<0.000), and the equation Y=1.029X4-8.181 was used. The monthly average air pressure 2 months prior(b=-2.418, β=-1.619, P<0.05)and the monthly average relative humidity(b=-0.739, β=-1.201, P<0.05)significantly negatively predicted the mosquito density. Notably, the regression equation used was y=2526.170+(-2.418)X9+(-0.739)X13. The exposure-response analysis revealed that the density of mosquitoes was not linearly related to the average monthly air pressure, rainfall, and duration of sunshine. Conclusions The dominant mosquito species around the BCIA was Cx. pipiens pallens, Ae. albopictus, and Ae. vexans. During the control of Cx. pipiens pallens, the impact on other mosquito species must also be considered. Environmental management, breeding sites, and scientific use of pesticides should be prioritized based on the activity periods of mosquitoes. Further studies on mosquitoes and meteorological factors should be conducted to provide references and novel avenues for mosquito control.
8.Colorimetric Detection of Sodium Dodecyl Benzene Sulfonate Based on Silver Phosphate/Nickel Hydroxystannate with Oxidase-like Activity
Qin HE ; Zhen-Bo YUAN ; Qi ZHANG ; Li-Li DU ; Bao-Jun HUANG ; Wei-Wei HE
Chinese Journal of Analytical Chemistry 2025;53(10):1654-1663
A highly efficient oxidase-mimetic silver phosphate/nickel hydroxystannate(Ag3PO4/NiSn(OH)6)composite was synthesized via a precipitation method using nickel hydroxystannate(NiSn(OH)6)as the support.The abundant hydroxyl groups(—OH)on NiSn(OH)6 not only provided nucleation sites for Ag3PO4 nanoparticles but also improved their dispersion and overall material stability.Based on oxidase-like activity of Ag3PO4/NiSn(OH)6 and inhibitory effect of sodium dodecylbenzenesulfonate(SDBS)on this catalytic activity,a novel colorimetric sensing method for SDBS detection was developed.Under optimized experimental conditions,the method exhibited a linear range of 3.69-42.7 μmol/L,with a detection limit of 0.135 μmol/L(S/N=3).The regression equation was ΔA652=0.01125C(μmol/L)+0.1498,with a correlation coefficient(R2)of 0.992.Practical application in dishwashing liquid analysis achieved satisfactory recoveries of 96.9%-106.4%,demonstrating the method's reliability for real sample detection.
9.Creation and Exploration of the"Organized Fill-in-the-Blank Format"Disci-pline Construction Model for Forensic Medicine in the New Era
Zhi-Wen WEI ; Hong-Xing WANG ; Jun-Hong SUN ; Hao-Liang FAN ; Hong-Liang SU ; Le-Le WANG ; Wen-Ting HE ; Zhe CHEN ; Jie ZHANG ; Xiang-Jie GUO ; Ji LI ; Geng-Qian ZHANG ; Xin-Hua LIANG ; Jiang-Wei YAN ; Qiang-Qiang ZHANG ; Cai-Rong GAO ; Ying-Yuan WANG ; Hong-Wei WANG ; Jun XIE ; Bo-Feng ZHU ; Ke-Ming YUN
Journal of Forensic Medicine 2025;41(1):25-29
Forensic medicine has been designated as a first-level discipline,presenting new opportunities and challenges for the development of forensic medicine.Since the 1980s,the establishment of foren-sic medicine discipline and the cultivation of high-level forensic talents have become hot topics in the development of forensic medicine in China.Since the 13th Five-Year Plan,the forensic team of Shanxi Medical University has been aiming at the forefront,proposing the development goals of"Five First-class"and the discipline development path"Six Major Achievements".It has selected benchmark disci-plines,identified gaps in disciplinary development,unified thoughts,formulated completion timelines,concentrated superior resources,assigned tasks to individuals,and created an"Organized Fill-in-the-Blank Format"forensic medicine discipline construction model with the characteristics of the new era.The construction model of forensic medicine has achieved good results in the goals,discipline frame-work,scientific research,talent cultivation,discipline team and platform construction,forming a rela-tively complete discipline construction and management system,and accumulating valuable experience for the construction of first-level discipline and high-level talent cultivation of forensic medicine.
10.Detection of Ketamine and Norketamine Using an Aptamer-Functionalized Gra-phene Oxide Fluorescent Sensor
Li-Xia WEI ; Bo LIU ; Xiao-Yuan YANG ; Xi ZHANG ; Yi-Feng LAN ; Chao ZHANG ; Juan JIA ; Dan ZHANG ; Zhi-Wen WEI ; Ke-Ming YUN ; Zhe CHEN
Journal of Forensic Medicine 2025;41(4):326-339
Objective To construct an aptamer-functionalized carboxylated graphene oxide(CGO)fluo-rescent sensor to achieve highly sensitive and specific detection of ketamine(KET)and its metabolite norketamine(NK)using an aptamer capable of simultaneously recognizing KET and NK.Methods A specific aptamer for simultaneous recognition of KET and NK was screened using graphene oxide-sys-tematic evolution of ligand by exponential enrichment(GO-SELEX)and molecular docking tech-niques.The aptamer,labeled with Cy5 fluorescence,was chemically conjugated to CGO to construct an aptamer-functionalized CGO fluorescent sensor.By optimizing detection conditions,including the mass concentration of CGO,aptamer concentration,reaction temperature,and incubation time,quantita-tive analysis of the target analytes was achieved using the ratio of fluorescence intensity changes be-fore and after target addition.The stability of the sensor in biological matrices was evaluated by moni-toring fluorescence intensity changes over incubation time in blank blood and urine,in comparison with the traditional physical adsorption-based CGO fluorescent sensor.Spiked recovery experiments in blank blood and urine were conducted to compare performance with that of HPLC-MS/MS.Results A specific aptamer A5 was selected and chemically conjugated with CGO to construct the aptamer-functionalized CGO fluorescent sensor.Under optimized conditions,the proposed fluorescent sensor ex-hibited a linear detection range of 1.0-5.0 ng/mL for KET,with a limit of detection(LOD)of 0.86 ng/mL;while for NK,the linear detection range was 1.0-5.0 ng/mL,with an LOD of 0.70 ng/mL.Com-pared with the CGO fluorescent sensor constructed via physical adsorption,this sensor demonstrated greater stability in blood and urine.The spiked recovery rates of KET and NK in blank blood and urine ranged from 81.50%to 110.03%,exhibiting detection performance comparable to that of HPLC-MS/MS.Conclusion The aptamer screening method offers a novel approach for selecting aptamers tar-geting drugs and their metabolites.The constructed aptamer-functionalized CGO fluorescent sensor pro-vides an efficient and reliable strategy for the high-performance detection of KET and NK.


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