1.Intervention Mechanism of Guizhi Fulingwan in Delaying Colitis-associated Colon Cancer via Modulating and Restoring MDSCs and Reshaping Immune Microenvironment
Yanwei HAO ; Chunrun LI ; Zhengwu QU ; Junmei TANG ; Jing GUO ; Yi ZHANG ; Fengming YOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):185-194
ObjectiveThis paper aims to investigate the efficacy and related actions of Guizhi Fulingwan in intervening in the mice with colitis-associated colon cancer (CAC) based on the immunosuppressive microenvironment associated with myeloid-derived suppressor cells (MDSCs). MethodsSixty male C57BL/6 mice were randomly assigned to a blank group, a model group, an aspirin group (0.04 g·kg-1), and low-, medium-, and high-dose Guizhi Fulingwan groups (4.87, 9.75, and 19.50 g·kg-1), with ten mice per group. The CAC mouse model was established via combined induction of azoxymethane (AOM)/dextran sulphate sodium (DSS). Drug intervention commenced in week five, with continuous intragastric administration for nine weeks. The food intake, body weight, fecal characteristics, and haematochezia were observed and recorded, and disease activity index (DAI) scores were calculated according to scoring criteria. Hematoxylin and eosin (HE) staining was used to observe the histopathological changes in the colon tissues of the mice. Immunohistochemistry was used to determine proliferating cell nuclear antigen-67 (Ki67) expression in the colon tissues, and enzyme-linked immunosorbent assay (ELISA) was used to detect the contents of interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α (TNF-α) in the serum of the mice. Flow cytometry was employed to determine the proportion levels of MDSCs, CD4+ T cells, and CD8+ T cells in the spleen tissues of the mice. The mRNA expressions of MDSC-associated effector molecules, including arginase 1 (Arg1) and inducible nitric oxide synthase (iNOS), were detected by real-time quantitative polymerase chain reaction (Real-time PCR). After that, an in vitro co-culture model of MDSCs and CD8+ T cells was established, and drug-containing serum of Guizhi Fulingwan was used for intervention. The Flow cytometry was employed to assess the effects of drug-containing serum of Guizhi Fulingwan with different concentrations on the levels of reactive oxygen species (ROS) and iNOS in MDSCs and the proliferation of CD8+ T cells. The levels of granzyme B (GZMB) and interferon-γ (IFN-γ) in cell supernatant were detected by ELISA. ResultsCompared with those in the control group, the mice in the model group exhibited significantly reduced body weight, elevated DAI scores, shortened colon length (P<0.01), increased number of tumors and Ki67 expression (P<0.01), and significantly elevated contents of IL-6, IL-1β, and TNF-α in the serum (P<0.01). Significant increases in the number of MDSCs were observed in mouse spleens, alongside marked reductions in the levels of CD4+ T and CD8+ T cells (P<0.01). Furthermore, the mRNA expressions of MDSC function-associated effector molecules Arg1 and iNOS were significantly upregulated (P<0.01). Compared with those in the model group, the mice in the middle-dose Guizhi Fulingwan group exhibited increased body weight and significantly decreased DAI scores (P<0.05, P<0.01). The mice in the middle- and high-dose Guizhi Fulingwan groups exhibited significantly improved colon shortening, significantly decreased number of tumors and Ki67 expression (P<0.05, P<0.01), and significantly decreased contents of IL-6, IL-1β, and TNF-α in the serum (P<0.05, P<0.01). Furthermore, administration of Guizhi Fulingwan markedly reduced MDSC infiltration in the spleen of the mice, with different degrees of increase in the levels of both CD4+ T and CD8+ T cells (P<0.05, P<0.01), alongside significant decreases in the mRNA expressions of Arg1 and iNOS (P<0.05, P<0.01). In vitro cell co-culture shows that administration of drug-containing serum of Guizhi Fulingwan significantly decreases the activity levels of ROS and iNOS in MDSCs and promotes the proliferation of CD8+ T cells and the secretion of GZMB and IFN-γ (P<0.05, P<0.01). ConclusionGuizhi Fulingwan can reduce pro-inflammatory cytokine secretion and inhibit tumor proliferation in the colon tissues of CAC mice. Its potential mechanism may involve reducing MDSC infiltration, enhancing effector T cells, particularly CD8+ T cell response, and improving the tumor immunosuppressive microenvironment.
2.Zhuluan Decoction Ameliorates Premature Ovarian Insufficiency by Inhibiting Excessive Autophagy of KGN Through Regulation of PI3K/Akt/mTOR Pathway
Yao CHEN ; Sainan TIAN ; Jing ZENG ; Xingxing YI ; Wen'e LIU ; Lei LEI ; Li TANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):89-98
ObjectiveTo elucidate the underlying mechanism through which Zhuluan decoction suppresses excessive autophagy in human ovarian granulosa cells (KGN) and ameliorates premature ovarian insufficiency (POI) via the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway. MethodsThe optimal concentration of cyclophosphamide for inducing a POI model in KGN cells was identified via the cell counting kit-8 (CCK-8) assay. Subsequently, the impacts of varying concentrations of Zhuluan decoction-containing serum on the viability of the KGN cell model were assessed. After the optimal drug concentration was determined, KGN cells were categorized into the following groups: blank control (20% blank serum), model (20% blank serum + 5 μmol·L-1 cyclophosphamide), Zhuluan decoction-containing serum (20% Zhuluan decoction-containing serum + 5 μmol·L-1 cyclophosphamide), autophagy inhibitor (20% blank serum + 5 μmol·L-1 cyclophosphamide + 20 μmol·L-1 chloroquine phosphate), autophagy inhibitor + Zhuluan decoction-containing serum (20% Zhuluan decoction-containing serum + 5 μmol·L-1 cyclophosphamide + 20 μmol·L-1 chloroquine phosphate), and estradiol valerate (20% estradiol valerate-containing serum + 5 μmol·L-1 cyclophosphamide). Following 48 hours of incubation, flow cytometry was utilized to measure the apoptosis rate of KGN cells in each group. Western blotting was employed to quantify the protein levels of PI3K, phosphorylated (p)-Akt, Akt, p-mTOR, and mTOR, along with the expression levels of autophagy-related proteins such as Beclin1, autophagy-related 5 homolog (ATG5), and microtubule-associated protein 1 light chain 3 (LC3), in each group. Additionally, monodansylcadaverine (MDC) staining was performed to evaluate the extent of autophagy in each group. ResultsIncubation of KGN cells with 5 μmol·L-1 cyclophosphamide for 48 h successfully established a POI model, marked by a significant inhibition of KGN cell proliferation. Notably, the inhibitory effect of cyclophosphamide on KGN cell proliferation exhibited a positive correlation with its concentration. Zhuluan decoction-containing serum at 20% and 30% promoted cell proliferation and mitigated the inhibitory effect of cyclophosphamide on KGN cell proliferation, with comparable therapeutic efficacy observed at both concentrations. Compared with the blank control group, the model group displayed an elevated apoptosis rate (P<0.01), reduced protein levels of PI3K, p-Akt, and p-mTOR (P<0.01), increased protein levels of Beclin1, LC3, and ATG5 (P<0.01), no significant alterations in the protein levels of Akt and mTOR, and an enhanced MDC autophagy fluorescence intensity (P<0.01). In comparison to that the model group, the apoptosis rates in the blank control group, model group, Zhuluan decoction-containing serum group, autophagy inhibitor group, autophagy inhibitor + Zhuluan decoction-containing serum group, and estradiol valerate group all reduced (P<0.05, P<0.01), with the most pronounced reduction observed in the autophagy inhibitor + Zhuluan decoction-containing serum group. The protein levels of PI3K, p-Akt, and p-mTOR were higher in other groups than in the model group (P<0.05, P<0.01), being the highest in the autophagy inhibitor + Zhuluan decoctio-containing serum group (P<0.01). The protein levels of Beclin1 and ATG5 were lower in other groups than in the model group (P<0.05, P<0.01). The expression level of LC3 declined in the Zhuluan decoction-containing serum group and the estradiol valerate group (P<0.05, P<0.01), while it decreased without statistical significance in the autophagy inhibitor group and the autophagy inhibitor + Zhuluan decoction-containing serum group. ConclusionZhuluan decoction may activate the PI3K/Akt/mTOR pathway to inhibit excessive autophagy and counteract the detrimental effects of cyclophosphamide on the KGN cell model, thus managing POI.
3.Zhuluan Decoction Ameliorates Premature Ovarian Insufficiency by Inhibiting Excessive Autophagy of KGN Through Regulation of PI3K/Akt/mTOR Pathway
Yao CHEN ; Sainan TIAN ; Jing ZENG ; Xingxing YI ; Wen'e LIU ; Lei LEI ; Li TANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):89-98
ObjectiveTo elucidate the underlying mechanism through which Zhuluan decoction suppresses excessive autophagy in human ovarian granulosa cells (KGN) and ameliorates premature ovarian insufficiency (POI) via the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway. MethodsThe optimal concentration of cyclophosphamide for inducing a POI model in KGN cells was identified via the cell counting kit-8 (CCK-8) assay. Subsequently, the impacts of varying concentrations of Zhuluan decoction-containing serum on the viability of the KGN cell model were assessed. After the optimal drug concentration was determined, KGN cells were categorized into the following groups: blank control (20% blank serum), model (20% blank serum + 5 μmol·L-1 cyclophosphamide), Zhuluan decoction-containing serum (20% Zhuluan decoction-containing serum + 5 μmol·L-1 cyclophosphamide), autophagy inhibitor (20% blank serum + 5 μmol·L-1 cyclophosphamide + 20 μmol·L-1 chloroquine phosphate), autophagy inhibitor + Zhuluan decoction-containing serum (20% Zhuluan decoction-containing serum + 5 μmol·L-1 cyclophosphamide + 20 μmol·L-1 chloroquine phosphate), and estradiol valerate (20% estradiol valerate-containing serum + 5 μmol·L-1 cyclophosphamide). Following 48 hours of incubation, flow cytometry was utilized to measure the apoptosis rate of KGN cells in each group. Western blotting was employed to quantify the protein levels of PI3K, phosphorylated (p)-Akt, Akt, p-mTOR, and mTOR, along with the expression levels of autophagy-related proteins such as Beclin1, autophagy-related 5 homolog (ATG5), and microtubule-associated protein 1 light chain 3 (LC3), in each group. Additionally, monodansylcadaverine (MDC) staining was performed to evaluate the extent of autophagy in each group. ResultsIncubation of KGN cells with 5 μmol·L-1 cyclophosphamide for 48 h successfully established a POI model, marked by a significant inhibition of KGN cell proliferation. Notably, the inhibitory effect of cyclophosphamide on KGN cell proliferation exhibited a positive correlation with its concentration. Zhuluan decoction-containing serum at 20% and 30% promoted cell proliferation and mitigated the inhibitory effect of cyclophosphamide on KGN cell proliferation, with comparable therapeutic efficacy observed at both concentrations. Compared with the blank control group, the model group displayed an elevated apoptosis rate (P<0.01), reduced protein levels of PI3K, p-Akt, and p-mTOR (P<0.01), increased protein levels of Beclin1, LC3, and ATG5 (P<0.01), no significant alterations in the protein levels of Akt and mTOR, and an enhanced MDC autophagy fluorescence intensity (P<0.01). In comparison to that the model group, the apoptosis rates in the blank control group, model group, Zhuluan decoction-containing serum group, autophagy inhibitor group, autophagy inhibitor + Zhuluan decoction-containing serum group, and estradiol valerate group all reduced (P<0.05, P<0.01), with the most pronounced reduction observed in the autophagy inhibitor + Zhuluan decoction-containing serum group. The protein levels of PI3K, p-Akt, and p-mTOR were higher in other groups than in the model group (P<0.05, P<0.01), being the highest in the autophagy inhibitor + Zhuluan decoctio-containing serum group (P<0.01). The protein levels of Beclin1 and ATG5 were lower in other groups than in the model group (P<0.05, P<0.01). The expression level of LC3 declined in the Zhuluan decoction-containing serum group and the estradiol valerate group (P<0.05, P<0.01), while it decreased without statistical significance in the autophagy inhibitor group and the autophagy inhibitor + Zhuluan decoction-containing serum group. ConclusionZhuluan decoction may activate the PI3K/Akt/mTOR pathway to inhibit excessive autophagy and counteract the detrimental effects of cyclophosphamide on the KGN cell model, thus managing POI.
4.Research on The Genealogical Inference Efficiency of High-density SNPs
Jing LI ; Yi-Jie SUN ; Wen-Ting ZHAO ; Zi-Chen TANG ; Jing LIU ; Cai-Xia LI
Progress in Biochemistry and Biophysics 2026;53(3):740-753
ObjectiveThis study aims to explore the potential of different orders of magnitude single-nucleotide polymorphism (SNP) locus combinations for predicting distant kinship relationships. A high-density SNP locus set was constructed, and a comprehensive assessment of its inference capability was conducted. MethodsFirstly, we selected three commercial chip panels, CGA (Chinese genotyping array, Illumina), GSA (Global screening array, Illumina), Affy (23MF_V2 high-density SNP array, Affymetrix) and merged them after quality control, forming a high-density SNP locus panel(1 180 k). Secondly, we selected 161 samples and collected their peripheral blood samples by using whole-genome sequencing technology. Within this sample population, the levels of kinship relationships fully covered the range from level 1 to level 9, and the number of kinship pairs at each level was consistently maintained at over 50 pairs. From 161 samples data of whole-genome sequencing, the 1 180 k locus set was extracted, which is referred to as the high-density SNP locus set in the following text. The kinship inference was conducted using the identity-by-descent (IBD) algorithm with the selected optimal parameters. To comprehensively evaluate the performance of the high-density SNP locus set in kinship inference, we compared it with the three commercial chip panels, the intersection of these three chip loci, and the control sets constructed by randomly reducing the number of the high-density SNP locus set. Based on the changes in the IBD lengths, as well as the dynamic trends in prediction accuracy, we conducted a scientific assessment of the kinship inference capability of the high-density SNP locus set. ResultsAfter screening, a set of 1 184 334 autosomal SNPs was obtained. During the process of screening the optimal IBD length threshold, the result revealed that 0 cM, 1 cM, and 2 cM all demonstrated good applicability. However, to avoid the issue of a large amount of redundant information caused by setting a too low IBD length threshold, this study ultimately selected 2 cM as the optimal threshold. Compared with the average results of three chip panels, the high-density SNP locus set increased the total IBD length and the average IBD length across levels 1-9; the accuracy of the confidence interval for level 8 was 70.97%, which represented a 3.50% improvement; the average confidence interval accuracy for levels 1-8 was 91.39%, representing a 1.00% increase; and the false negative rates at levels 8 and 9 were reduced by 2.42% and 6.76%, respectively. The system efficacy of the high-density SNP locus set for kinship inference of first to eighth degree relationships reached 98.91%. Through random reduction of the high-density SNP locus set results, it is found that increasing the number of SNPs with the panel, the detection efficiency of IBD length showed a significant upward trend. At the same time, the overall trend in the accuracy of kinship relationship prediction as well as the confidence interval accuracy also indicated that both metrics steadily increased with the addition of more loci. ConclusionThe results show that the high-density SNPs panel significantly enhances the efficacy of distant kinship inference, accurately covering kinship degrees, with the average confidence interval accuracy for first to eighth degree relationships stably above 90%. The study finds that increasing the number of SNPs panel can improve the ability to predict distant kinship.
5.Surveillance of soil-transmitted nematodiasis in Hubei Province from 2016 to 2023
Juan ZHANG ; Wen LIN ; Hong ZHU ; Lun WAN ; Jia YI ; Dongni WU ; Jing XIA ; Li TANG
Chinese Journal of Schistosomiasis Control 2026;38(3):287-292
Objective To investigate the prevalence of soil-transmitted nematode human infections in Hubei Province from 2016 to 2023, so as to provide insights into formulation of soil-transmitted nematodiasis control strategies and evaluation of soil-transmitted nematodiasis control effectiveness. Methods There were 10 surveillance points established in Hubei Province for soil-transmitted nematodiasis each year from 2016 to 2023. The surveillance points were divided into eastern, western, southern, northern and central regions according to geographical locations, and one administrative village was sampled from a township in each region as a surveillance point. Then, at least 200 permanent residents at ages of over 3 years were randomly sampled at each point, with at least 1 000 individuals sampled. Soil-transmitted nematode infection was detected using a modified Kato-Katz thick smear method (two slides for one stool sample), and Enterobius vermicularis infection was additionally detected with a cellophane tape test among children at ages of 3 to 9 years. The prevalence of soil-transmitted nematode human infections was descriptively analyzed. Results A total of 80 547 individuals underwent surveillance for soil-transmitted nematodiasis in Hubei Province from 2016 to 2023, and a total of 262 individuals were detected with soil-transmitted nematode infections, including two cases with mixed infections, yielding an overall prevalence rate of 0.33% (262/80 547). The prevalence rates of hookworm, Ascaris lumbricoides, Trichuris trichiura, and E. vermicularis infections were 0.02% (20/80 547), 0.07% (55/80 547), 0.02% (15/80 547) and 0.22% (174/80 547), respectively, appearing a parasite species-specific prevalence rate (χ2 = 250.235, P < 0.05), and the prevalence rates of soil-transmitted nematode infections were 0.50%(51/10271),0.37%(34/9120),0.39%(39/10099),0.40%(41/10168), 0.36% (37/10 234), 0.45% (46/10 211), 0.03% (3/10 238) and 0.11% (11/10 206) in Hubei Province from 2016 to 2023, respectively, showing a year-specific prevalence rate (χ2 = 60.589, P < 0.05). Soil-transmitted nematode infections were detected across 36 surveillance points, and there was a significant difference in the prevalence of soil-transmitted nematode infections among surveillance points (0.02% to 3.30%, P < 0.001). In addition, the differences in prevalence rates of human soil-transmitted nematode infection among different age groups (χ2 = 531.091, P < 0.05), occupations (P < 0.001), and education levels (P < 0.001) were all statistically significant. Among these, the children at ages of 3 to 9 years old (1.40%, 174/12 461), nursery-school children (1.69%, 90/5 312) and preschool-aged children (1.56%, 93/5 969) were relatively high. Conclusions The overall prevalence of soil-transmitted nematodiasis was low in Hubei Province from 2016 to 2023; however, the prevalence varied across regions and among populations, with E. vermicularis as the predominant species of soil-transmitted nematode. Soil-transmitted nematodiasis control should be given a high priority in key areas in Hubei Province, and preschool children, primary school students, and individuals with a low educational level are recommended as key target populations for health education and interventions.
6.Mechanistic investigation of Fuzheng Hefu Zhiyang Formula in alleviating psoriasis inflammatory microenvironment via P38/Erk/NF-κB signaling pathway
Yi-jing LIAO ; Yan-jie LIU ; Yue LU ; Bin TANG ; Jun-hong ZHANG ; Jing-jie YU ; Hao DENG ; Ling HAN ; Chuan-jian LU ; Hai-ming CHEN
Chinese Traditional Patent Medicine 2025;47(8):2550-2558
AIM To investigate the effect of Fuzheng Hefu Zhiyang Formula(FZHFZY)on psoriasis-like skin lesions and immune regulation in mice.METHODS In the in vivo experiment,30 BALB/c mice were randomly divided into the blank group,the model group,the dexamethasone group(1.5 g/kg of compound dexamethasone acetate cream),and the low-dose(2.5 g/kg)and high-dose(5 g/kg)FZHFZY groups,with six mice in each group.The experiment groups were treated with respective FZHFZY and dexamethasone,and the other groups were given normal saline for 10 consecutive days,during which psoriatic skin lesions were induced with imiquimod cream for 7 consecutive days.The mice had their area and severity of psoriasis assessed by PASI score;their histological changes of skin lesions.observed with Hematoxylin-eosin(HE)staining;their F4/80 ratio of skin lesions observed with immunohistochemical(IHC)staining;their protein expressions of P38,p-P38,Erk,p-Erk,P65 and p-P65 detected by Western blot;and their mRNA expressions of tumor necrosis factor-α(TNF-α),IL-17,IL-23 and IL-1β detected by RT-qPCR.In the in vitro research,the cultured RAW264.7 cells were divided into the blank group,the LPS group,and the FZHFZY groups(1 200,600,300,150 μg/mL).The cells had their protein expressions of P38,p-P38,Erk,p-Erk,P65 and p-P65 detected with Western blot;and their mRNA expressions of IL-6,TNF-α,IL-23 and IL-8 detected by RT-qPCR.RESULTS The in vivo experiment showed that compared to the model group,the FZHFZY groups demonstrated decreased PASI score(P<0.01);improved epidermal thickening and parakeratosis of skin lesions as revealed by HE staining result and increased expression of F4/80 in IHC staining sections;decreased protein expression ratios of p-P38/P38,p-ERK/Erk and p-P65/P65 in skin(P<0.05,P<0.01);and reduced mRNA expressions of TNF-α,IL-17,IL-23 and IL-1β in the skin(P<0.01).FZHFZY(0~2 400 μg/mL)showed no significant cytotoxicity towards RAW264.7 cells in vitro(P>0.05).Compared to those of the LPS group,the cells exposed to FZHFZ at concentrations of 1 200 and 600 μg/mL demonstrated decreased protein expression ratios of p-P38/P38,p-ERK/Erk,and p-P65/P65(P<0.05,P<0.01);and significantly decreased mRNA expressions of TNF-α,IL-17,IL-23 and IL-1β(P<0.01).CONCLUSION FZHFZY alleviates imiquimod-induced psoriatic lesions in mice and suppresses inflammatory response in LPS-stimulated RAW264.7 cells by inhibiting P38/Erk/NF-κB signaling pathway.
7.Study on the Theory of"Any Deficiency of Spleen and Stomach Being A Disease of the Blood"
Journal of Zhejiang Chinese Medical University 2025;49(7):923-927,934
[Objective]To investigate LI Dongyuan's theory of"any deficiency of spleen and stomach is a disease of the blood"and provide reference for modern clinical practice.[Methods]Taking Discussion on Internal and External Injuries,Treatise on Spleen and Stomach,Orchid Chamber of Secrets and Invention of Medicine as references,it starts from the connotation of"any deficiency of spleen and stomach is a disease of the blood",and explores the etiology and pathogenesis of the diseases,the theoretical construction and the clinical application respectively.[Results]Taking Inner Canon of Yellow Emperor and Zhongjing's academic thought as the source,absorbing CHENGs' and ZHU Xi's ideas of science.Being inspired by his teacher ZHANG Yuansu,and driven by his own clinical practice,LI Dongyuan put forward the academic thought of"any deficiency of spleen and stomach is a disease of the blood".From the Treatise on Spleen and Stomach-Treatise on the Prosperity and Decline of Spleen and Stomach,"any deficiency of spleen and stomach is a disease of the blood",which is a pathological state caused by the deficiency of spleen and stomach Qi and the weakness of elevation due to the disorders of emotions and moods,overwork and overdose,and uncontrolled diets,there fore causes insufficient blood production and too much blood consumption,and then leads to the deficiency of blood and Yin,and the essence of which is the disordered ascending,descending,floating and sinking of the human body's Qi.In treatment,LI Dongyuan always based on regulating blood,to restore the body's Qi,and harmonize Qi and blood as the goal,with"blood replenishment must have Yang Qi,Qi and blood double tonic""rising Yang must be in response to the time of the day,and make good use of the wind herbs""relieving heat must protect the Yin and blood,moderation in attacking and destroying".[Conclusion]Exploring this theory can help to view the relationship between Qi and blood from a new perspective,better grasp LI Dongyuan's academic thinking,and provide reference for the treatment of spleen and stomach diseases and blood diseases.
8.Determination of 71 Kinds of Per-and Polyfluoroalkyl Substances in Eckloniae Thallus by QuEChERS-Liquid Chromatography-Tandem Mass Spectrometry
Dan-Rui TANG ; Jing SUN ; Ling CAO ; Dan-Yi JIANG ; Hai-Wei SHI ; Lu-Lu WU
Chinese Journal of Analytical Chemistry 2025;53(7):1177-1185,中插9-中插25
A QuEChERS cleanup-ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was established for simultaneous determination of traditional and emerging 71 kinds of per-and polyfluoroalkyl substances(PFAS)in Chinese herbal medicine Eckloniae Thallus.The extraction solvent employed here was 0.2%formic acid-acetonitrile with sodium chloride-anhydrous sodium sulfate(3:1,m/m)as salting-out agent,and the purification was carried out using graphitized carbon black and octadecyl-bonded silica as adsorbents.The sample separation was carried out on a Horizon C18 chromatographic column(150 mm×2.1 mm,1.6 μm)by gradient elution with 2.5 mmol/L ammonium acetate aqueous solution(containing 2.5 mmol/L acetic acid)and acetonitrile as the mobile phases.The flow rate was set at 0.4 mL/min,with an injection volume of 5 μL.The chromatographic separation of each target compound and internal standard substance could be achieved within 15 min.An electrospray ion source was utilized for simultaneous scanning of positive and negative ions under the scheduled multiple reaction monitoring(sMRM)mode,and the internal standard method was used for quantitative analysis.The results of method validations showed that the 71 kinds of PFAS had good linearity,with a correlation coefficient(r)greater than 0.997,spiked recoveries of 76.2%?122.0%and relative standard deviations(RSDs)of 5.5%?14.8%.The limits of detection were in the range of 0.3-60 ng/kg and the limits of quantification were in the range of 0.8-199 ng/kg.The developed method was used for detection of PFAS in 14 batches of Eckloniae Thallus samples,and a total of 19 kinds of PFAS were detected.The findings indicated that PFAS residues were commonly present in Eckloniae Thallus,with perfluorocarboxylic acids(PFCAs)exhibiting the highest concentrations.The method was simple,robust and efficient in preparation,sensitive in detection with high throughput,and suitable for monitoring residual PFAS compounds,including both ionic and neutral in food-medicine homology samples such as Eckloniae Thallus.
9.Progress in Methods for Electrochemical Detection of Thrombin
Di WU ; Xi-Yao ZHANG ; Jing-Jing XU ; Yi-Ting CHEN ; Wen-Qi TANG ; Wen-Hui XU ; Song-Min CHEN ; Qiong HU ; Li NIU
Chinese Journal of Analytical Chemistry 2025;53(9):1403-1410
As a serine protease,thrombin can convert soluble fibrinogen into insoluble fibrin and plays a pivotal role in the coagulation cascade.Therefore,the accurate quantitative assay of thrombin levels is of great value in the evaluation of coagulation function,clinical screening and prognostic monitoring of coagulation-related diseases,and screening of drugs for targeted therapy.Existing methods for thrombin detection can be divided into two categories,e.g.,the assay of concentration levels using nucleic acid aptamers as the affinity elements and the assay of activity levels based on the hydrolytic cleavage of substrate peptides.In recent years,electrochemical biosensors have attracted much attention in thrombin detection due to high sensitivity,high selectivity,simple instrument,fast response,and good portability.In this review,the latest research progress in methods for electrochemical detection of thrombin was summarized,focusing on the detection principles and the applied signal amplification strategies of related electrochemical biosensors.In addition,the challenges with respect to the practical use of electrochemical thrombin biosensors and the prospects were discussed.
10.The role and research progress of m6A modification in sepsis and its induced multi-organ dysfunction disease
Lu-lu ZHANG ; Rui GONG ; Jin-yi ZHAO ; Fei MU ; Yan-ping YIN ; Wang-ting LI ; Ling-ling ZHENG ; Yu-ping TANG ; Jing-wen WANG
Chinese Pharmacological Bulletin 2025;41(3):421-427
Sepsis is a life-threatening organ dysfunction disease caused by a dysregulated host response to infection.It has com-plex pathophysiological changes,and in severe cases,it can rap-idly develop into septic shock and multiple organ dysfunction or multiple organ failure.At present,the pathological mechanism of sepsis and its induced organ dysfunction is complex and the in-fluencing factors are numerous.So far,there is still a lack of specific and effective treatment strategies.RNA modify-N6-methyladenosine(m6 A)is one of the most common post-tran-scriptional modifications on eukaryotic RNAs.It is involved in the regulation of the occurrence and development of a variety of inflammatory diseases,including sepsis,and even multiple organ dysfunction induced by sepsis by affecting the metabolism of RNAs.It includes cardiac dysfunction,acute lung injury(ALI)and acute kidney injury(AKI).Therefore,this article will dis-cuss the effect of m6A modification on the function of immune cells,and its important role in sepsis and its induced multiple or-gan dysfunction diseases by regulating inflammatory signals,py-roptosis,mitochondrial damage and ferroptosis.This will provide new therapeutic targets and strategies for the clinical prevention and treatment of sepsis and its induced multiple organ dysfunc-tion diseases.

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