1.Establishment and Evaluation of an Oxidative Stress Model of Atopic Dermatitis Induced by 2,4-dinitrofluorobenzene
Chang LIU ; Xuesong XIANG ; Huihuang HE ; Xiaoqing CHEN ; Wenhong QIU
Laboratory Animal and Comparative Medicine 2026;46(1):46-54
Objective To establish an oxidative stress mouse model of atopic dermatitis (AD) by applying 2,4-dinitrofluorobenzene (DNFB) to the back and post-auricular skin of KM mice, and to evaluate the regulatory role of the RAGE-NLRP3 axis (receptor for advanced glycation end products-NOD-like receptor family, pyrin domain containing 3 axis) in AD-related oxidative stress, thereby providing a potential therapeutic target for AD treatment. Methods Twenty SPF-grade female KM mice were randomly divided into a control group (Control group) and an experimental group (DNFB group), with 10 mice in each group. Mice in the Control group were treated with an acetone-olive oil vehicle (acetone: olive oil = 3:1) on their back and post-auricular skin. Mice in the DNFB group were treated with 0.5% DNFB (prepared by adding 0.5 g DNFB per 100 mL of acetone-olive oil vehicle) on the same areas, once daily for 14 consecutive days. The severity of skin lesions was scored on days 2, 4, 6, 9, 12, and 14 of treatment. On day 14, scratching behavior and ear thickness were evaluated. Ear swelling was evaluated on the final day by measuring bilateral ear thickness three times with a vernier caliper; the three measurements were averaged. HE staining was used to observe morphological and structural changes of cells in the back skin tissues. The mRNA and protein expression levels of RAGE (receptor for advanced glycation end products) in skin tissues were detected by quantitative real-time PCR, Western blot, and immunohistochemical staining. The mRNA expression levels of oxidative stress-related molecules, including NLRP3 (NOD-like receptor family, pyrin domain containing 3), caspase-1 (cysteine-dependent aspartate-specific protease 1), and IL-1β (Interleukin-1β), were detected by quantitative real-time PCR. Results On day 14, the back skin lesion scores of the Control group and DNFB group were (0.20±0.42) and (9.93±1.30) (P<0.000 1), respectively. Scratching behavior scores were (5.00±2.05) and (49.26±8.49) episodes, respectively (P<0.000 1), and ear thicknesses were (213.00±11.87) μm and (765.93±140.47) μm (P<0.000 1), respectively. The DNFB group exhibited marked skin dryness, desquamation, and thickening. HE staining results showed that skin inflammation was obvious in the DNFB group, consistent with the pathological features of AD. Quantitative real-time PCR and Western blot results showed that compared with the Control group, the mRNA expression level of RAGE in skin tissues of the DNFB group was significantly increased (P<0.05), and the protein expression level of RAGE was also significantly increased (P<0.01). Immunohistochemical staining results showed that compared with the Control group, skin tissue sections of the DNFB group exhibited thickened stratum corneum and fibrotic proliferation of fibroblasts in the interstitium under microscopic observation, with a significant increase in RAGE protein expression in the skin tissues (P<0.01). Quantitative real-time PCR results showed that the mRNA expression levels of NLRP3, caspase-1, and IL-1β in skin tissues of the DNFB group were all significantly increased (P<0.01). Conclusion The AD mouse oxidative stress model has been successfully established by topical DNFB application. RAGE may promote the development of AD by regulating the NLRP3 inflammasome and IL-1β release, forming an oxidative-inflammatory cascade, suggesting that it could be a potential therapeutic target for AD.
2.Study on the apoptosis-inducing effect of esculetin on acute myeloid leukemia HL-60 cells via regulating the AKT/SKP2/MTH1 pathway
Weihua SONG ; Fuying CHU ; Wei XIE ; Jinliang CHEN ; Ping ZHAO ; Hong QIU ; Jian TAO ; Xiang CHEN
China Pharmacy 2026;37(1):36-41
OBJECTIVE To investigate the apoptosis-inducing effect of esculetin (Esc) on acute myeloid leukemia (AML) HL-60 cells by regulating the protein kinase B (AKT)/S-phase kinase-associated protein 2 (SKP2)/MutT homolog 1 (MTH1) pathway. METHODS AML HL-60 cells were randomly divided into control group (routine culture), Esc low-concentration group (L-Esc group, 25 μmol/L Esc), Esc medium-concentration group (M-Esc group, 50 μmol/L Esc), Esc high-concentration group (H-Esc group, 100 μmol/L Esc), and high-concentration of Esc+ SC79 (AKT agonist) group (100 μmol/L Esc+5 μmol/L SC79). Cell proliferation in each group was detected by MTT assay and colony formation assay. The level of reactive oxygen species (ROS) in cells was measured by using the CM-H2DCFDA fluorescent probe. Cell apoptosis was analyzed by flow cytometry. Western blot assay was performed to detect the expression levels of apoptosis-related proteins [B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), cleaved caspase-3], AKT/SKP2/MTH1 pathway-related proteins (p-AKT, AKT, SKP2, MTH1), along with the upstream and downstream proteins of AKT phosphatidylinositol 3-kinase (PI3K), cyclin-dependent kinase inhibitor 1 (P21) and cyclin-dependent kinase inhibitor 1B (P27). RESULTS Compared with control group, the cell viability, colony number, and the phosphorylation levels of AKT and PI3K proteins as well as protein expressions of SKP2, MTH1 and Bcl-2 were significantly decreased (P<0.05), while ROS level, apoptosis rate, and the expression levels of Bax, cleaved caspase-3, P21 and P27 proteins were significantly increased (P<0.05). Moreover, the effects of Esc exhibited concentration-dependence (P<0.05). Compared with H-Esc group, above indexes of high-concentration of Esc+ SC79 group were reversed significantly (P<0.05). CONCLUSIONS Esc may promote massive ROS production and induce activation of apoptosis in HL-60 cells by inhibiting the AKT/SKP2/MTH1 pathway, thus inhibiting the proliferation of HL-60 cells.
3.Development of brush ionization probe mass spectrometry for convenient on-site detection of traditional Chinese medicine
Junxian WU ; Chaofa WEI ; Ceyu MIAO ; Jiaquan XU ; Xiang LI ; Li ZHOU ; Shuanglong WANG ; Liping KANG ; Zidong QIU
Science of Traditional Chinese Medicine 2026;4(1):81-86
Objective: To develop a convenient, direct, and highly sensitive method for screening trace chemical additives in complex Chinese patent medicines, thereby addressing core technological bottlenecks in pharmaceutical analysis and quality control. Methods: A brush ionization probe device was independently designed and constructed, and an efficient detection method was established through systematic optimization of key parameters. Twenty-three Chinese patent medicine samples, representing 6 dosage forms (capsules, tablets, pills, granules, powders, and liquid preparations), were analyzed using 10 common chemical additives as target analytes. Results: All samples were successfully analyzed without complex pretreatment, and 5 chemical additives were detected in 7 Chinese patent medicines. The brush ionization probe device exhibited cost-effectiveness (~0.2 USD per probe), operational simplicity, rapid analysis (~10s per sample), high efficiency, and minimal reagent consumption (~10 μL per sample). Conclusion: This advancement is expected to provide an innovative scientific tool for improving the generality and convenience of on-site quality control, while promoting technological progress in disciplines such as pharmacology and traditional Chinese medicine.
4.Value of deep-learning combined with 0.55 T MRI in evaluating pulmonary tumors
Xiang WANG ; Wei LI ; Jia LIU ; Shuai MA ; Jianxing QIU
Chinese Journal of Radiology 2025;59(7):765-770
Objective:To investigate the value of 0.55 T MRI scanner using deep-learning (DL) reconstruction in evaluating pulmonary tumors.Methods:The study was a cross-sectional study. Sixty-one patients with pulmonary tumors on CT images were prospectively collected from May to September 2024 in Peking University First Hospital, including 37 males and 24 females, and aged 46?89 (68±9) years old. All patients underwent lung scan on a 0.55 T MRI, using diffusion weighted imaging(DWI) sequence with b-values of 0 and 800 s/mm 2. According to whether DL reconstruction was used and the number of acquisitions, they were divided into DWI-DL 5∶30 group (DL, number of averages=10, acquisition time=5 min 30 s),DWI-DL 3∶22 group (DL, number of averages=5, acquisition time=3 min 22 s), and DWI-C group (GRAPPA, number of averages=10, acquisition time=5 min 30 s). The obtained images were evaluated subjectively (Likert score) and objectively [signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR)]. Meanwhile the apparent diffusion coefficient (ADC) value of the tumors was measured. Friedman nonparametric test was used for comparison among the three groups and Bonferroni method was used for pairwise comparison. Results:The subjective scores, SNR, and CNR were significantly different among DWI-DL 5∶30 group, DWI-DL 3∶22 group, and DWI-C group( χ 2=9.69,87.56,88.62, P=0.008,<0.001,<0.001). Bonferroni method results showed that the subjective scores, SNR, and CNR of DWI-DL 5∶30 group were higher than those of DWI-DL 3∶22 group and DWI-C group ( P<0.05); However, the subjective scores, SNR, and CNR did not significantly differ between DWI-DL 3∶22 group and DWI-C group ( P>0.05). The ADC values of the tumors were not significantly different among DWI-DL 5∶30 group, DWI-DL 3∶22 group, and DWI-C group (χ 2=5.95, P=0.510). Conclusion:The DWI reconstructed using DL has better or similar image quality to conventional DWI in evaluating pulmonary tumors and significantly reduces scanning time, which has certain clinical application value.
5.Development and challenges of mass spectrometry database for traditional Chinese medicine: A review
Yijun WANG ; Zhiming YANG ; Junxian WU ; Xiaoli MA ; Li ZHOU ; Xiang LI ; Baiping MA ; Zidong QIU ; Liping KANG
Science of Traditional Chinese Medicine 2025;3(3):210-221
Accurate characterization of the chemical composition of complex traditional Chinese medicine (TCM) is an essential foundation for the modern scientific interpretation of TCM principles. Mass spectrometry is the most dominant technique in current research on the material basis of TCM, offering the highest sensitivity and the richest information provision. Establishing mass spectrometry databases represents the most effective approach to facilitating the structural analysis of TCM chemical components. This paper systematically searches and reviews literature published from January 2005 to January 2025 through online databases such as China National Knowledge Infrastructure, PubMed, and Web of Science, using “mass spectrometry database” and “traditional Chinese medicine” as keywords. It reviews the current status of seven TCM chemical component mass spectrometry databases and seven natural product mass spectrometry databases. The key advancements of these mass spectrometry databases for natural products are summarized, detailing their characteristics, search methodologies, included information, and data sources. Additionally, challenges related to data quality, standardization, timely updates, database interaction, retrieval functionality, and data sharing and security are discussed in depth. Furthermore, the paper explores prospective development directions for TCM mass spectrometry databases, emphasizing the importance of open data sharing, technological innovation, and data security. Through this analysis, the paper aims to offer theoretical guidance and practical recommendations for the precise identification of TCM components, as well as for the construction and application of these databases.
6.Value of deep-learning combined with 0.55 T MRI in evaluating pulmonary tumors
Xiang WANG ; Wei LI ; Jia LIU ; Shuai MA ; Jianxing QIU
Chinese Journal of Radiology 2025;59(7):765-770
Objective:To investigate the value of 0.55 T MRI scanner using deep-learning (DL) reconstruction in evaluating pulmonary tumors.Methods:The study was a cross-sectional study. Sixty-one patients with pulmonary tumors on CT images were prospectively collected from May to September 2024 in Peking University First Hospital, including 37 males and 24 females, and aged 46?89 (68±9) years old. All patients underwent lung scan on a 0.55 T MRI, using diffusion weighted imaging(DWI) sequence with b-values of 0 and 800 s/mm 2. According to whether DL reconstruction was used and the number of acquisitions, they were divided into DWI-DL 5∶30 group (DL, number of averages=10, acquisition time=5 min 30 s),DWI-DL 3∶22 group (DL, number of averages=5, acquisition time=3 min 22 s), and DWI-C group (GRAPPA, number of averages=10, acquisition time=5 min 30 s). The obtained images were evaluated subjectively (Likert score) and objectively [signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR)]. Meanwhile the apparent diffusion coefficient (ADC) value of the tumors was measured. Friedman nonparametric test was used for comparison among the three groups and Bonferroni method was used for pairwise comparison. Results:The subjective scores, SNR, and CNR were significantly different among DWI-DL 5∶30 group, DWI-DL 3∶22 group, and DWI-C group( χ 2=9.69,87.56,88.62, P=0.008,<0.001,<0.001). Bonferroni method results showed that the subjective scores, SNR, and CNR of DWI-DL 5∶30 group were higher than those of DWI-DL 3∶22 group and DWI-C group ( P<0.05); However, the subjective scores, SNR, and CNR did not significantly differ between DWI-DL 3∶22 group and DWI-C group ( P>0.05). The ADC values of the tumors were not significantly different among DWI-DL 5∶30 group, DWI-DL 3∶22 group, and DWI-C group (χ 2=5.95, P=0.510). Conclusion:The DWI reconstructed using DL has better or similar image quality to conventional DWI in evaluating pulmonary tumors and significantly reduces scanning time, which has certain clinical application value.
7.Construction and identification of recombinant fowl adenovirus 4 expressing Cap protein of goose astrovirus virus genotype 2
Xingyu LI ; Yan LI ; Panpan YANG ; Junjie LIU ; Mengjia XIANG ; Yutao ZHU ; Luyao QIU ; Qilong QIAO ; Boshun ZHANG ; Dexin BU ; Chenghao HAN ; Chunmei YU ; Yanfang CONG ; Zeng WANG ; Jianli LI ; Baiyu WANG ; Jun ZHAO
Chinese Journal of Veterinary Science 2025;45(3):443-448,513
To construct a recombinant fowl adenovirus 4(FAdV-4)expressing the Cap protein of goose astrovirus genotype 2(GoAstV-2),the expression cassette of Cap gene was inserted into the natural 1 966 bp deletion region of the FAdV-4 genome in the infectious clone p15A-cm-FAdV4-HNJZ.The resulted recombinant plasmid p15A-cm-FAdV4-HNJZ-Cap/GoAstV-2 was linearized with restriction enzyme and transfected into chicken hepatoma cell line(LMH)to rescue the recombinant FAdV-4 expressing the Cap protein of GoAstV-2,rF Ad V4-Cap/GoAstV-2.After 15 passages in LMH cells,the recombinant rFAdV4-Cap/GoAstV-2 was identified by PCR using primers flanking the insertion site of the Cap gene expression cassette and using viral genome DNA extracted from rFAdV4-Cap/GoAstV-2 infected LMH cells as template.LMH cells were in-fected with 15th passage rFAdV4-Cap/GoAstV-2 and indirect immunofluorescence was performed with a polyclonal antibody against Cap protein as the primary antibody.Western blot was carried out with lysates of rFAdV4-Cap/GoAstV-2 infected LMH cells.The in vitro replication dynamic of the 15th passage of the rFAdV4-Cap/GoAstV-2 was also investigated in LMH cells.The results demonstrated that the Cap gene of GoAstV-2 was presented in the genome of the recombinant vi-rus rF AdV4-Cap/Go Ast V-2,and could be expressed stably.The prepared recombinant virus in this study will lay a foundation for developing inactivated bivalent vaccine candidate against co-in-fection of FAdV-4 and GoAstV-2 in goose.
8.Preparation of sustained-release microspheres of ginsenosides
Xiang-ying LIU ; Ru-nan HE ; Gao QIU ; Zong-hao LIN ; Yu-hai GUO ; Chu-qin YU
Chinese Traditional Patent Medicine 2025;47(4):1097-1104
AIM To prepare the sustained-release microspheres of ginsenosides.METHODS The sustained-release microspheres were prepared by SPG membrane emulsification technology with poly(lactic-co-glycolic acid)(PLGA)as a shell carrier.With PLGA concentration,feed rate and Span 60 concentration as influencing factors,comprehensive score for appearance,drug loading and encapsulation efficiency as an evaluation indice,the preparation process was optimized by response surface method.The morphology of sustained-release microspheres was observed,after which the particle size,drug loading and encapsulation efficiency were determined,and the in vitro drug release was investigated.RESULTS The optimal conditons were determined to be 45 s for agitation time of primary emulsion,74.68 mg/mL for PLGA concentration,11%for feed rate,and 4.18 mg/mL for Span 60 concentration,the comprehensive score was 74.98.The round sustained-release microspheres demonstrated the average particle size of 4.33 μm,drug loading of(8.24±0.13)%,and encapsulation efficiency of(74.94±1.17)%,respectively.At 336 h,ginsenosides Rg1,Rb1,Rb2 displayed the accumulative release rates of 84.12%,78.04%,65.88%,respectively.CONCLUSION This reasonable and feasible method can be used for the preparation of sustained-release microspheres of ginsenosides with good appearance and high drug loading,which can provide references for the preparation of other water-soluble drug microspheres and solution of microsphere collapse problem.
9.Expert consensus on the model informed precision dosing of tacroli-mus in patients receiving anti-rejection therapy
Bing CHEN ; Xiaocong ZUO ; Xingang LI ; Dewei SHANG ; Peijun ZHOU ; Junjie DING ; Xiaoq-iang XIANG ; Xiaoyan QIU ; Zhuo WANG ; Xiaoyu LI ; Yi ZHANG ; Wei ZHAO ; Yuzhu WANG ; Jianjun GAO ; Zheng JI-AO
Chinese Journal of Clinical Pharmacology and Therapeutics 2025;30(4):433-445
There is significant inter-individual variation of pharmacokinetics and pharmacody-namics in patients receiving tacrolimus(TAC)for an-ti-rejection therapy,which cause the rejection or toxic action.Based on results of therapeutic drug monitoring and pathophysiological index of trans-plant patients,the individualized dosing regimen can be designed and adjusted by using model in-formed precision dosing(MIPD).The patients'clini-cal outcome can be improved.In the consensus,the different methods of MIPD used for patients re-ceived TAC for anti-rejection therapy were intro-duced,which can be used for the designing and ad-justing doing regimen,predicting adverse drug reac-tion,improving medication adherence and econom-ics during therapy.
10.Surveillance and analysis of drug resistance molecular markers in Plasmodium vivax of imported cases in Chongqing
Yao XIANG ; Yan TAN ; Fei LUO ; Jiaojiao CAI ; Zhifeng LI ; Jingru XU ; Jingfu QIU
Chinese Journal of Zoonoses 2025;41(7):726-734
This research aimed to comprehensively understand the prevalence of mutation in drug-resistant molecular markers of imported Plasmodium vivax in Chongqing,the Pvmdr1,Pvdhps,Pvdhfr,Pvcrt-o and Pvk12 genes of Plasmodium vivax were systematically analyzed.Blood samples were collected from imported Plasmodium vivax-infected patients in Chongqing between 2011 and 2022.The Pvmdr1,Pvdhps,Pvdhfr,Pvcrt-o,and Pvk12 genes were amplified and then sequenced to precisely evaluate gene mutations.Bioinformatics methods were employed to conduct in depth analysis of the mutation prevalence.Regarding the Pvdhfr gene,mutations at codons 50,57,58,61,99,117,and 199 were detected in 2.9%,23.5%,76.4%,23.53%,2.9%,82.3%,and 5.88%of the samples,respectively.The double-mutant haplotype S58R/S117N was the most prevalent,accounting for 50%,followed by the quadruple-mutant haplotype F57L/S58R/T61M/S117T,which accounted for 11.76%.Among the four types of tandem-repeat variations of Pvdhfr,the wild-type was the most common,and the insertion type was a novel discovery in this study.For the Pvdhps gene,the prevalence among mutation genotypes was relatively low.The single-mutant genotype was dominant,constituting 27.03%.The prevalence of Pvmdr1 mutations at codons 958 and 1076 was 100%and 89.19%,respectively.Among the 37 successfully sequenced samples,K10 insertion was detected in only 8 cases(22.22%).Notably,no non-synonymous mutations of Pvk12 were identified in this study.The cases in this study were imported from various countries of origin.Novel tandem-repeat variation tyres of Pvdhfr and new mutation sites of Pvdhps were identifide,thus enriching the mutation information of imported Plasmodium vivax resistance molecular markers in China.

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