1.Da Chaihutang for Treatment of Sepsis with Yang Syndrome:A Randomized Controlled Trial
Na HUANG ; Guangmei CHEN ; Xingyu KAO ; Zhen YANG ; Weixian XU ; Kang YUAN ; Junna LEI ; Jingli CHEN ; Mingfeng HE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):55-63
ObjectiveTo explore the clinical efficacy and safety of Da Chaihutang (DCH) for the treatment of sepsis with Yang syndrome. MethodsA total of 70 patients suffering from sepsis with Yang syndrome were randomly divided into an observation group and a control group, with 35 cases in each group. They both received standard Western medicine treatment. The observation group was additionally given a dose of DCH, which was boiled into 100 mL and taken twice. The control group was additionally given an equal volume and dosage of warm water. The intervention lasted for three days. The 28-day all-cause mortality and the changes in the following indicators before and after intervention were compared between the two groups, including sequential organ failure assessment (SOFA), acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score,white blood cell (WBC),the percentage of neutrophils (NEU%),C-reactive protein (CRP),procalcitonin (PCT),alanine transaminase (ALT),aspartate transaminase (AST),total bilirubin (TBil),creatinine (Cr),blood urea nitrogen (BUN),acute gastrointestinal injury (AGI) grade,gastrointestinal dysfunction score (GDS),serum intestinal fatty acid-binding protein (iFABP), citrulline (CR),platelet (PLT),prothrombin time(PT),activated partial thromboplastin time (APTT),fibrinogen (Fib),international normalized ratio (INR),and D-dimer (D-D). ResultsThere was no significant difference between the two groups regarding 28-day all-cause mortality. After the intervention,SOFA,WBC,PCT,and Cr were significantly decreased, and PLT was significantly increased in the control group (P<0.05). SOFA,APACHE Ⅱ,NEU%,CRP,PCT,ALT,AST,Cr,BUN,AGI grade,GDS,and serum iFABP and CR were significantly improved in the observation group (P<0.05). After the intervention,APACHE Ⅱ,PCT,AGI grade,GDS,and serum iFABP in the observation group were significantly lower than those in the control group ,while CR and PLT were higher (P<0.05,P<0.01). There were significant differences regarding the gap of SOFA,APACHE Ⅱ,AST,TBil,AGI grade,GDS,iFABP,CR, and PLT between the two groups (P<0.05,P<0.01). There were slight differences regarding PT,APTT,Fib,INR,and D-D between the two groups,which were in the clinical normal range. ConclusionOn the basis of Western medicine, DCH helped to reduce sepsis severity and improved multiple organ dysfunction with high clinical efficacy and safety, but further research on its impact on the prognosis of patients with sepsis is still required.
2.Current status of eating behaviors and its predictive role in overweight and obese of adolescents
Chinese Journal of School Health 2025;46(1):53-57
Objective:
To explore the current status and influencing factors of eating behaviors in adolescents, so as to provide a theoretical foundation for health promotion education among adolescents.
Methods:
Based on the database from Survey of Chinese Family Health Index (2021), by a random number table method, 1 065 teenagers were selected from the provincial capitals of 22 provinces and 5 autonomous regions in China, as well as 4 municipalities directly under the central government. A general characteristic questionnaire, Patient Health Questionnaire-9 (PHQ-9), Short Form of the Family Health Scale (FHS-SF), 10-item Short Version of the Big Five Personality(BFP-10), Content-based Media Exposure Scale (CM-E) and Sakata Eating Behavior Scale Short Form(EBS-SF) were used to collect information. Multivariate stepwise linear regression analysis was employed to identify and analyze related factors of eating behaviors among adolescents. Receiver operating characteristic was used to validate the predictive ability of the EBS-SF score for overweight and obesity among adolescents.
Results:
The average scores of BFI-10,C-ME, FHS-SF, PHQ-9 and EBS-SF were (33.08±4.64)(19.20±4.55)(38.48±6.65)(6.09±5.63)(16.75±4.36), respectively. Multivariate linear regression showed that family type (other types), agreeableness, conscientiousness, family health and depression were the main related factors of EBS-SF scores among adolescents( B =2.61,-0.42,0.20,-0.11,0.23, P <0.05).The analysis of receiver operating characteristic curve showed that the EBS-SF scores had a good ability in predicting obesity among male adolescents ( AUC= 0.73, P <0.01).
Conclusions
Family type, big five personality, family health,depression are the related factors of eating behaviors among adolescents. EBS-SF scores are predictive of obesity in adolescents, which would provide a new perspective for promoting healthy eating habits among adolescents.
3.Influencing factors of neonatal red blood cell transfusion: a retrospective analysis
Na ZHOU ; Xin HE ; Yu SI ; Chen HOU ; Jialu CHEN ; Zhaohui TANG
Chinese Journal of Blood Transfusion 2025;38(3):375-381
[Objective] To analyze the effects of different factors and red blood cell transfusion thresholds on the efficacy of neonatal red blood cell (RBC) transfusion, in order to provide more references for neonatal transfusions to better achieve rational and effective blood use. [Methods] A retrospective collection of data from 282 neonates who received RBC transfusions at our hospital from 2022 to 2023 was conducted, including birth weight, gestational age, number of blood transfusions, length of hospital stay, assisted ventilation during RBC transfusion, and laboratory test results before and after transfusion. SPSS software was used for statistical analysis to comprehensively analyze the impact of different factors on the efficacy of RBC transfusion in neonates. [Results] The results showed that the gestational age and weight of newborns at birth were negatively correlated with their length of hospital stay and the number of RBC transfusions during hospitalization. Newborns with younger gestational age and lower weight had longer hospital stays and more RBC transfusions during hospitalization. After administering RBCs according to the standard of 15 mL/kg, there was a statistically significant difference in the efficacy of RBC transfusion at different transfusion thresholds. In non-critical situations, RBC transfusions were ineffective when the pre-transfusion hemoglobin (Hb) level was >120 g/L. When the pre-transfusion Hb level was ≤70 g/L, RBC transfusions achieved higher efficacy in both critical and non-critical situations. [Conclusion] In critical situations, the group with pre-transfusion Hb values ≤ 70 g/L has the best RBC transfusion effect, while in non-critical situations, the group with pre-transfusion Hb levels between 81 and 90 g/L has the best RBC transfusion effect. Overall, the efficacy of RBC transfusion in non-critical situations is higher than that in critical situations.
4.Oxylipidomics Combined with Transcriptomics Reveals Mechanism of Jianpi Huogu Prescription in Treating Steroid-induced Osteonecrosis of Femoral Head in Rats
Lili WANG ; Qun LI ; Zhixing HU ; Qianqian YAN ; Liting XU ; Xiaoxiao WANG ; Chunyan ZHU ; Yanqiong ZHANG ; Weiheng CHEN ; Haijun HE ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):190-199
ObjectiveTo unveil the mechanism of Jianpi Huogu prescription (JPHGP) in ameliorating the dyslipidemia of steroid-induced osteonecrosis of the femur head (SONFH) by oxylipidomics combined with transcriptomics. MethodsSixty SD rats were assigned into normal, model, low-, medium-, and high-dose (2.5, 5, 10 g·kg-1, respectively) JPHGP, and Jiangushengwan (1.53 g·kg-1) groups. Lipopolysaccharide was injected into the tail vein at a dose of 20 μg·kg-1 on days 1 and 2, and methylprednisolone sodium succinate was injected at a dose of 40 mg·kg-1 into the buttock muscle on days 3 to 5. The normal group received an equal volume of normal saline. Drug administration by gavage began 4 weeks after the last injection, and samples were taken after administration for 8 weeks. Hematoxylin-eosin staining was conducted to reveal the histopathological changes of the femoral head, and the number of adipocytes, the rate of empty bone lacunae, and the trabecular area were calculated. Micro-computed tomography was used for revealing the histological and histomorphometrical changes of the femoral head. Enzyme-linked immunosorbent assay was employed to measure the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), apolipoprotein A1 (ApoA1), and apolipoprotein B (ApoB). At the same time, the femoral head was collected for oxylipidomic and transcriptomic detection. The differential metabolites and differential genes were enriched and analyzed, and the target genes regulating lipid metabolism were predicted. The predicted target proteins were further verified by molecular docking, immunohistochemistry, and Western blot. ResultsCompared with the normal group, the model group showcased thinning of the femoral head, trabecular fracture, karyopyknosis, subchondral cystic degeneration, increases in the number of adipocytes and the rate of empty bone lacunae (P<0.01), a reduction in the trabecular area (P<0.01), decreases in BMD, Tb.Th, Tb.N, and BV/TV, and increases in Tb.Sp and BS/BV (P<0.01). Compared with the model group, the JPHGP groups showed no obvious thinning of the femoral head or subchondroidal cystic degeneration. The high- and medium-dose JPHGP groups presented declines in the number of adipocytes and the rate of empty bone lacunae, an increase in the trabecular area (P<0.05, P<0.01), rises in BMD, Tb.Th, Tb.N, and BV/TV, and decreases in Tb.Sp and BS/BV (P<0.05, P<0.01). Compared with the normal group, the model group showcased raised serum levels of TG, TC, LDL, and ApoB and lowered serum levels of HDL and ApoA1 (P<0.01). Compared with the model group, the JPHGP groups had lowered serum levels of TG, TC, LDL, and ApoB (P<0.05, P<0.01) and a risen serum level of ApoA1 (P<0.05, P<0.01). Moreover, the serum level of HDL in the high-dose JPHGP group increased (P<0.01). A total of 19 different metabolites of disease set and drug set were screened out by oxylipidomics of the femoral head, and 119 core genes with restored expression were detected by transcriptomics. The enriched pathways were mainly concentrated in inflammation, lipids, apoptosis, and osteoclast differentiation. Molecular docking, immunohistochemistry, and Western blot results showed that compared with the normal group, the model group displayed increased content of 5-lipoxygenase (5-LO) and peroxisome proliferator-activated receptor γ (PPARγ) in the femoral head (P<0.01). Compared with the model group, medium- and high-dose JPHGP reduced the content of 5-LO and PPARγ (P<0.05, P<0.01). ConclusionJPHGP can restore the levels of oxidized lipid metabolites by regulating the 5-LO-PPARγ axis to treat SONFH in rats. Relevant studies provide experimental evidence for the efficacy mechanism of JPHGP in the treatment of SONFH.
5.Oxylipidomics Combined with Transcriptomics Reveals Mechanism of Jianpi Huogu Prescription in Treating Steroid-induced Osteonecrosis of Femoral Head in Rats
Lili WANG ; Qun LI ; Zhixing HU ; Qianqian YAN ; Liting XU ; Xiaoxiao WANG ; Chunyan ZHU ; Yanqiong ZHANG ; Weiheng CHEN ; Haijun HE ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):190-199
ObjectiveTo unveil the mechanism of Jianpi Huogu prescription (JPHGP) in ameliorating the dyslipidemia of steroid-induced osteonecrosis of the femur head (SONFH) by oxylipidomics combined with transcriptomics. MethodsSixty SD rats were assigned into normal, model, low-, medium-, and high-dose (2.5, 5, 10 g·kg-1, respectively) JPHGP, and Jiangushengwan (1.53 g·kg-1) groups. Lipopolysaccharide was injected into the tail vein at a dose of 20 μg·kg-1 on days 1 and 2, and methylprednisolone sodium succinate was injected at a dose of 40 mg·kg-1 into the buttock muscle on days 3 to 5. The normal group received an equal volume of normal saline. Drug administration by gavage began 4 weeks after the last injection, and samples were taken after administration for 8 weeks. Hematoxylin-eosin staining was conducted to reveal the histopathological changes of the femoral head, and the number of adipocytes, the rate of empty bone lacunae, and the trabecular area were calculated. Micro-computed tomography was used for revealing the histological and histomorphometrical changes of the femoral head. Enzyme-linked immunosorbent assay was employed to measure the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), apolipoprotein A1 (ApoA1), and apolipoprotein B (ApoB). At the same time, the femoral head was collected for oxylipidomic and transcriptomic detection. The differential metabolites and differential genes were enriched and analyzed, and the target genes regulating lipid metabolism were predicted. The predicted target proteins were further verified by molecular docking, immunohistochemistry, and Western blot. ResultsCompared with the normal group, the model group showcased thinning of the femoral head, trabecular fracture, karyopyknosis, subchondral cystic degeneration, increases in the number of adipocytes and the rate of empty bone lacunae (P<0.01), a reduction in the trabecular area (P<0.01), decreases in BMD, Tb.Th, Tb.N, and BV/TV, and increases in Tb.Sp and BS/BV (P<0.01). Compared with the model group, the JPHGP groups showed no obvious thinning of the femoral head or subchondroidal cystic degeneration. The high- and medium-dose JPHGP groups presented declines in the number of adipocytes and the rate of empty bone lacunae, an increase in the trabecular area (P<0.05, P<0.01), rises in BMD, Tb.Th, Tb.N, and BV/TV, and decreases in Tb.Sp and BS/BV (P<0.05, P<0.01). Compared with the normal group, the model group showcased raised serum levels of TG, TC, LDL, and ApoB and lowered serum levels of HDL and ApoA1 (P<0.01). Compared with the model group, the JPHGP groups had lowered serum levels of TG, TC, LDL, and ApoB (P<0.05, P<0.01) and a risen serum level of ApoA1 (P<0.05, P<0.01). Moreover, the serum level of HDL in the high-dose JPHGP group increased (P<0.01). A total of 19 different metabolites of disease set and drug set were screened out by oxylipidomics of the femoral head, and 119 core genes with restored expression were detected by transcriptomics. The enriched pathways were mainly concentrated in inflammation, lipids, apoptosis, and osteoclast differentiation. Molecular docking, immunohistochemistry, and Western blot results showed that compared with the normal group, the model group displayed increased content of 5-lipoxygenase (5-LO) and peroxisome proliferator-activated receptor γ (PPARγ) in the femoral head (P<0.01). Compared with the model group, medium- and high-dose JPHGP reduced the content of 5-LO and PPARγ (P<0.05, P<0.01). ConclusionJPHGP can restore the levels of oxidized lipid metabolites by regulating the 5-LO-PPARγ axis to treat SONFH in rats. Relevant studies provide experimental evidence for the efficacy mechanism of JPHGP in the treatment of SONFH.
6.Molecular mechanisms and cancer-promoting roles of sodium arsenite in regulating FNDC3B expression in A549 cells
Jinyun JIANG ; He MA ; Na LIU ; Ruihuan ZHAO ; Chenglan JIANG ; Yuefeng HE
Journal of Environmental and Occupational Medicine 2025;42(6):691-698
Background Arsenic exposure has been demonstrated to induce apoptosis. The fibronectin type III structural domain 3B protein (FNDC3B) has been shown to promote cancer cell proliferation; however, its role in arsenic-induced apoptosis remains to be elucidated. Objective To investigate the effects of sodium arsenite (NaAsO2) and its metabolites on the expression of FNDC3B gene in A549 cells and to understand the function of FNDC3B gene in A549 cells. Methods (1) A549 cells were exposed to varying final concentrations of NaAsO2 and their optical density at 450 nm values were measured by Cell Counting Kit-8 (CCK-8) after 48 h. Survival curves were plotted, and a final exposure dose was selected according to the survival rate. Total protein and RNA were extracted by exposing A549 cells to high (30 µmol·L−1), medium (20 µmol·L−1), and low (10 µmol·L−1) NaAsO2 concentrations, high (30 µmol·L−1) monomethylarsinic acid (MMA), and high (30 µmol·L−1) dimethylarsinic acid for a period of 48 h. mRNA expression and the protein expression of the FNDC3B gene was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot (WB), while the protein ubiquitination expression of the FNDC3B gene was detected by co-immunoprecipitation (CO-IP) and WB assay. (2) Knockdown of FNDC3B gene expression was achieved in A549 cells by siRNA interference. The si-FNDC3B fragment was transfected in A549 cells for 48 h. The mRNA and protein expression of FNDC3B gene was then detected by qRT-PCR and WB assay. Cell viability was determined through CCK-8 assay. Hoechst 33342/propidium iodide (PI) double staining and JC-1 mitochondrial membrane potential assay were employed to detect both early and late apoptosis, while cleaved caspase3 protein and P53 signalling pathway related protein expressions were evaluated by WB. Results (1) The CCK-8 results demonstrated a decline in the viability of A549 cells with an increase in NaAsO2 concentration, with an inhibitory concentration at 50% of 38.12 µmol·L−1. The qRT-PCR results demonstrated that compared to the control group, varying concentrations of NaAsO₂ (10, 20, and 30 µmol·L⁻¹) significantly upregulated the mRNA expression of FNDC3B gene (P<0.01). In contrast, MMA and DMA showed no significant effect on FNDC3B mRNA expression (P>0.05). The WB analysis revealed that the protein expression of FNDC3B was reduced in the NaAsO₂-treated group compared to the control, accompanied by elevated ubiquitination levels of FNDC3B protein, particularly at the K48 ubiquitination site. MMA and DMA exhibited no impact on FNDC3B protein expression. (2) Following the specific knockdown of FNDC3B expression in A549 cells, the CCK-8 assay demonstrated a significant reduction in cell viability in the silenced FNDC3B group (si-FNDC3B) compared to the control group. The JC-1 assay demonstrated that the mitochondrial membrane potential was diminished in the si-FNDC3B group relative to the control group. The Hoechst 33342/PI staining assay revealed that the si-FNDC3B group exhibited a notable degree of apoptosis. The si-FNDC3B group also displayed substantial apoptosis. The WB analysis indicated that the relative expressions of cleaved caspase3, P53, MDM2, Bad, and Bax proteins were elevated in the si-FNDC3B group in comparison to the control group. Conclusion The presence of NaAsO2 is observed to promote the ubiquitination expression of the FNDC3B protein, which in turn reduces the expression of FNDC3B protein. However, the main metabolites DMA and MMA have no effect on the expression of FNDC3B. Furthermore, the silencing of FNDC3B is observed to inhibit the viability of A549 cells and promote apoptosis, a phenomenon related to the activation of P53 signaling pathway.
7.Network analysis of anxiety, depression and perceived stress with eating behaviors in adolescents
Chinese Journal of School Health 2025;46(6):821-826
Objective:
To explore the network structure of eating behaviors with anxiety, depression and perceived stress in adolescents, so as to provide a basis for effective prevention and intervention of eating behavior problems and negative emotions in adolescents.
Methods:
Based on the Psychology and Behavior Investigation of Chinese Residents (2021) database, the study was conducted among 3 087 adolescents. Sakata Eating Behavior Scale Short From(EBS-SF) was used to investigate their eating behaviors. The Patient Health Questionnaire-9(PHQ-9), Generalized Anxiety Disorder Scale-7 Item(GAD-7), and Perceived Stress Questionnaire-3 Item (PSQ-3) were used to evaluate their depression, anxiety and perceived stress. Network analysis method was applied to construct a network of eating behaviors and negative emotional symptoms among adolescents, so as to evaluate the centrality, bridge strength, stability and accuracy of each item.
Results:
The total scores of eating behaviors, depression,anxiety and stress perception in adolescents were 17.41±4.53,6.95±6.08,4.86±5.03,9.34±3.80,respectively. The symptom with the highest intensity and expected impact was "I am only satisfied when I buy more food than I need", with a node intensity and expected impact value of 4.37. The nodes Depression and Anxiety were the most closely connected(weight=0.87). There were no statistically significant differences in the network structure( M =0.13,0.11) and network connection strength(female and male:4.16,4.06, s =0.10;urban and rural areas:4.08,4.07, s =0.01) between different sexes and residents ( P >0.05).
Conclusion
The negative impact of comorbidities such as anxiety, depression, perceived stress and eating behaviors among adolescents can be reduced through targeted prevention and intervention of core symptoms and bridging symptoms.
8.Characteristics of HIV primary drug resistance and molecular transmission clusters in newly reported men who had sex with men in Taizhou City, Zhejiang Province
Shanling WANG ; Xuanhe WU ; Guixia LI ; Tingting WANG ; Yating WANG ; Tailin CHEN ; Weiwei SHEN ; Yali XIE ; Haijiang LIN ; Na HE ; Xiaoxiao CHEN
Shanghai Journal of Preventive Medicine 2025;37(6):496-502
ObjectivesTo investigate the molecular epidemiological characteristics of HIV-1 infection among men who had sex with men (MSM) in Taizhou City, Zhejiang Province, and to provide a scientific reference for acquired immune deficiency syndrome prevention and control efforts. MethodsThe research subjects were all newly reported MSM population in Taizhou City from 2020 to 2023. Blood samples without antiviral therapy were collected. The HIV-1 pol gene was amplified and sequenced, and the sequences were submitted to the Stanford University drug resistance database to identify the mutation sites and drug resistance. MEGA 11.0 software was used to analyze the nucleic acid sequences, construct phylogenetic tree, and calculate genetic distance of gene sequences. The molecular transmission network diagram of HIV-1 was constructed using Cytoscape_v3.10.1, and the influencing factors of network entry were analyzed by logistic regression. ResultsA total of 363 newly reported HIV-infected MSM patients were included, with a median age [M (P25, P75)] of 34 (26,47) years old. The majority had an educational level of junior high school or below (55.65%). A total of eight subtypes were found, mainly CRF07_BC and CRF01_AE. The primary drug resistance rate was 10.47% (38/363). The optimal molecular network gene distance was 0.019, with a network access rate of 42.70% (155/363), and a total of 47 molecular clusters were formed. Multivariate logistic analyses showed that compared with the CRF01_AE subtype, the clustering risk of CRF07_BC subtype was higher (OR=1.916, 95%CI: 1.191‒3.109), cases with drug resistance had a higher risk of cluster formation than those without drug resistance (OR=2.011, 95%CI: 1.006‒4.080), and recent infected patients had a lower risk of entering the largest molecular cluster than long-term infected patients (OR=0.376, 95%CI: 0.137‒0.928). ConclusionThe newly diagnosed infections among the MSM population are active in Taizhou City, Zhejiang Province, with a high level of primary drug resistance. Individuals carrying drug-resistant strains are more likely to cluster. Drug resistance monitoring should be strengthened to prevent further spread of drug-resistant strains in the network.
9.Efficacy of direct-acting antiviral agents combined regimens for hepatitis C virus with different genotypes in Dehong Prefecture, Yunnan Province from 2022 to 2024
Renhai TANG ; Yidan ZHAO ; Yuecheng YANG ; Runhua YE ; Lifen XIANG ; Xingmei FENG ; Qunbo ZHOU ; Yanfen CAO ; Na HE ; Yingying DING ; Song DUAN
Shanghai Journal of Preventive Medicine 2025;37(8):676-681
ObjectiveTo investigate the therapeutic effects of direct-acting antiviral agents (DAAs) combined regimens for hepatitis C virus (HCV) patients in Dehong Prefecture, Yunnan Province from 2022 to 2024, to analyze the characteristics of treatment failure patients, so as to provide a basis for discovering more effective treatment regimens in the future. MethodsData on HCV prevention and treatment in Dehong Prefecture was extracted from the China Disease Control and Prevention Information System. A total of 617 patients with HCV antiviral therapy were included, and the differences in variable characteristics among patients with different genotypes were analyzed using comparative statistical tests, including basic socio-demographic characteristics, biochemical testing indicators, and information on previous treatment and current treatment. In addition, the cure rate of HCV patients with diverse characteristics was compared, and the potential causes of treatment failure were explored simultaneously. ResultsThe cure rate of HCV was 96.8%, and statistically significant differences were observed in aspartate transaminase (AST) and alanine transaminase (ALT) levels, previous antiviral therapy history and initial treatment regimens among patients with different HCV genotypes (all P<0.05). Among the multi-type combination regimens, the cure rate of sofosbuvir (SOF)-containing regimens was 97.00%, that of velpatasvir (VEL)-containing regimens was 95.45%, and the cure rate of other treatment regimens, including the regimens with ribavirin (RIB) intervention, was 93.10%. Among the patients with treatment failure, 45.00% had genotype 3, 40.00% had abnormal abdominal ultrasound results, and all presented with elevated baseline AST test levels. ConclusionThe clinical treatment of HCV patients should consider the differences in genotype and biochemical test results. DAAs combined regimens for HCV have achieved a high cure rate in Dehong Prefecture and are applicable to HCV patients with diverse clinical characteristics, providing research evidence for wider application.
10.Untargeted Metabolomics Reveals Mechanism of Modified Sinisan in Ameliorating Anxiety-like Behaviors Induced by Chronic Restraint Stress in Mice
Jie ZHAO ; Zhengyu FANG ; He XIAO ; Na GUO ; Hongwei WU ; Hongjun YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):70-79
ObjectiveTo elucidate the potential mechanism of modified Sinisan (MSNS) in alleviating anxiety-like behaviors induced by chronic restraint stress (CRS) in mice at the metabolic level based on serum untargeted metabolomics and identify key metabolites and metabolic pathways regulated by MSNS. MethodsSeventy-two male C57BL/6 mice were randomly assigned into six groups: control, model, high-dose (2.4 g·kg-1) MSNS, medium-dose (1.2 g·kg-1) MSNS, low-dose (0.6 g·kg-1) MSNS, and positive control (fluoxetine, 2.6 mg·kg-1). Except the control group, the other groups were subjected to CRS for the modeling of anxiety. Mice were administrated with corresponding agents by gavage 2 h before daily restraint for 14 days. Anxiety-like behaviors were evaluated by the open field test (OFT), elevated plus maze (EPM) test, and light/dark box (LDB) test. Serum levels of corticotropin-releasing hormone (CRH), adrenocorticotrophic hormone (ACTH), and corticosterone (CORT) were measured via ELISA to assess stress levels. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was employed to detect 9 metabolites in the brain tissue and serum metabolites. Orthogonal partial least squares-discriminant analysis (OPLS-DA) was adopted to identify differential metabolites (VIP>1.0, P<0.05). MetaboAnalyst 5.0 was used for metabolic pathway enrichment analysis of the differential metabolites. ResultsCompared with the control group, the model group showed reductions in the central activity time and central distance in the OFT (P<0.05), the proportions of open-arm residence time and open-arm residence times in the EPM test (P<0.01), and the proportions of open box activity time and open box activity distance in the LDB test (P<0.05), which were increased in the medium- and high-dose MSNS groups compared with the model group (P<0.05). Compared with the control group, the model group showed elevated levels of CRH, ACTH, and CORT in the serum (P<0.01), and the elevations were diminished in the medium- and high-dose MSNS groups (P<0.05). UPLC-MS results indicated that compared with the control group, the model group presented declined DA, GABA, 5-HIAA, 5-HT, and 5-HT/Trp levels (P<0.05, P<0.01) and raised Glu, NE, Kyn, and Kyn/Trp levels (P<0.05). Compared with the model group, high-dose MSNS increased the GABA, 5-HIAA, and 5-HT/Trp levels (P<0.05) and lowered the Glu and Kyn/Trp levels (P<0.05). Untargeted metabolomics identified that 16 CRS-induced metabolic disturbances were reversed by MSNS. KEGG pathway analysis indicated that MSNS primarily modulated eight core pathways including alanine/aspartate/glutamate metabolism, butyrate metabolism, arginine-proline metabolism, TCA cycle, unsaturated fatty acid biosynthesis, and tryptophan metabolism. The mechanisms involved multidimensional biological processes, including neurotransmitter homeostasis regulation, TCA cycle energy metabolism optimization, and inflammatory response suppression. ConclusionMSNS alleviates CRS-induced anxiety-like behaviors in mice by mitigating hypothalamic-pituitary-adrenal axis hyperactivity, improving hippocampal neurotransmitter and tryptophan metabolic pathways, and regulating alanine/aspartate/glutamate metabolism, butyrate metabolism, arginine-proline metabolism, and TCA cycle.


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