1.Huangqi Jianzhongtang Regulates Polarization of Macrophages M1/M2 and Improves Fat Consumption in Cancer Cachexia Mice
Zhiyan FANG ; Haiyan ZHU ; Wenying HUAI ; Cong HUANG ; Ruocong YANG ; Haiyan YU ; Tiane ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):61-69
ObjectiveTo investigate the effects of Huangqi Jianzhongtang (HQJZ) on macrophage polarization and fat consumption in cancer cachexia (CC) mice. MethodsUltra-performance liquid chromatography-quadrupole/electrostatic field Orbitrap high-resolution mass spectrometry (UPLC-Q-Orbitrap HRMS) was used to control the quality of HQJZ. (1) In vitro experiment: HQJZ-containing serum was prepared, and the optimal concentration was determined by cytotoxicity assay. Mouse monocyte-derived macrophages (RAW264.7) were cultured and randomly divided into six groups, including a blank group, a classically activated macrophages (M1) group, an alternatively activated macrophages (M2) group, a HQJZ + blank group, a HQJZ+M1 group, and a HQJZ + M2 group. The relative expression of macrophage marker genes CD86, inducible nitric oxide synthase (iNOS), CD206, and arginase-1 (Arg1) was detected by real-time quantitative polymerase chain reaction (Real-time PCR ). (2) In vivo experiment: Thirty-two BALB/c mice were randomly divided into a control group, a model group, a medroxyprogesterone acetate (MPA) group, and a HQJZ group. Except for the control group, the other mice were injected with CT-26 colon cancer cells to establish a CC model. Mice in the MPA and HQJZ groups were given MPA (0.13 g·kg-1·d-1) or HQJZ (13.13 g·kg-1·d-1) by gavage, respectively, while mice in the control and model groups were given an equal volume of saline by gavage, with interventions continued for 10 d. Real-time PCR was used to detect the expression of macrophage markers (iNOS, Arg1, CD86, CD206) and fat browning-related genes uncoupling protein 1 (UCP1) and peroxisome proliferator-activated receptor γ (PPARγ) in epididymal adipose tissue. Western blot (WB) was used to detect protein expression levels of UCP1 and PPARγ. Micro-computed tomography (micro-CT) was used to measure residual fat volume, and hematoxylin-eosin (HE) staining was used to assess fat browning and calculate pathological scores. ResultsIn vitro, the dominant effective concentration of HQJZ-containing serum was 12.5%. Real-time PCR results showed that, compared with the blank group, Arg1 expression decreased in the HQJZ+blank group (P<0.05), CD206 showed a downward trend without statistical significance, while iNOS and CD86 expression were significantly increased (P<0.05). Compared with the M1 group, Arg1 and CD206 expression decreased in the HQJZ+M1 group (P<0.05). Compared with the M2 group, CD206 expression decreased in the HQJZ+M2 group (P<0.05), CD86 expression increased significantly (P<0.01). In vivo, Real-time PCR results showed that, compared with the control group, CD86 and CD206 expression levels were significantly increased in the model group (P<0.01). Compared with the model group, CD206 expression in the MPA group was significantly decreased (P<0.01). In the HQJZ group, CD206 was significantly decreased (P<0.01). WB results showed that, compared with the model group, protein expression of UCP1 and PPARγ was significantly reduced in the HQJZ group (P<0.05, P<0.01). micro-CT results showed that the total white fat volume in the HQJZ group was greater than that in the model group (P<0.05). HE staining results showed that pathological scores in the HQJZ group were lower than those in the model group (P<0.05). ConclusionHQJZ may inhibit white adipose tissue browning by promoting macrophage M1 polarization and suppressing M2 polarization, thereby delaying fat consumption in CC mice.
2.Comparative study on clinical characteristics of patients with retinal artery occlusion in different age groups
Chanjuan* HUANG ; Huhai* YUAN ; Haiyan XU ; Zirui PENG ; Pai LIU ; Rongdi YUAN
International Eye Science 2026;26(10):1690-1697
AIM: To examine differences in etiology, risk factors, gender, and acute-phase manifestations of retinal artery occlusion(RAO)patients in different age groups, highlighting age-related heterogeneity.
METHODS:This retrospective comparative cohort study enrolled patients with RAO. Patients were divided into <50 y group and ≥50 y group. Etiological classification was based on modified TOAST criteria. Comparisons assessed the prevalence of risk factors, etiological distribution, sex ratio, and, in cases of acute central retinal artery occlusion(CRAO), fundus characteristics and visual outcomes.
RESULTS:This study included a total of 145 RAO patients,which were divided into two groups based on age: <50 y group(15 females and 13 males)with the mean age of 36.64±9.72 y, and ≥50 y group(38 females and 79 males)with the mean age of 63.50±9.54 y. Hypertension(46.4% vs 85.5%)and diabetes(3.6% vs 27.4%)were less prevalent in <50 y group(both P<0.001). The <50 y group was characterized predominantly by stroke of other determined etiology(SOE, 35.7% vs 2.6%, P<0.001), whereas the ≥50 y group was dominated by small artery occlusion(SAO, 35.9%)and large artery atherosclerosis(LAA, 31.6%), the incidence rates were significantly higher in the ≥50 y group than in the <50 y group(both P<0.001). Females were more frequently represented in <50 y group(53.6% vs 32.5%, P=0.039). Multivariable analysis demonstrated that LAA was independently linked to older age and SOE to younger age. Boxcarring occurred exclusively in ≥50 y patients\〖39.5%(17/43)vs 0, P=0.011\〗.
CONCLUSION:The RAO patients<50 y predominantly present with non-atherosclerotic causes and a higher proportion of females, whereas patients≥50 y are mainly affected by atherosclerosis driven etiologies and show signs of reduced vascular compensatory capacity. These findings indicate that age-related differences in RAO should be taken into account when determining etiologic evaluation and management strategies.
3.Clinical Utility of Whole Exome Sequencing in the Diagnosis of Fetal Short Femur
Manqi CHEN ; Zewei HUANG ; Qi XU ; Shuning ZHANG ; Xiaojun CHEN ; Haiyan YAN ; Xiang LI ; Shuai FU
Journal of Sun Yat-sen University(Medical Sciences) 2026;47(2):368-378
ObjectiveTo enhance the genetic diagnostic yield for fetal short femur length (FL) and to evaluate the integration of whole exome sequencing (WES) with chromosomal abnormality detection in prenatal diagnosis. MethodsA total of 27 mid-to-late trimester fetuses diagnosed with short FL [FL < 5th percentile for gestational age or < -2 SD (Standard Deviation)] at the Prenatal Diagnosis And Medical Genetics Center,Shenshan Medical Center,Memorial Hospital of Sun Yat-Sen University between November 2023 and May 2025 were included. Amniotic fluid or umbilical cord blood samples were collected and subjected to chromosomal karyotyping (19cases), chromosomal microarray analysis (CMA) or low-coverage whole genome copy number variation(CNV-Seq) (27 cases), and trio-based whole exome sequencing (trio-WES) (27 cases). The pathogenicity of genetic variants were interpreted according to the American College of Medical Genetics and Genomics (ACMG) guidelines. ResultsTwenty-six genomic variants were detected in 17 of the 26 fetuses, including numerical chromosomal abnormalities (mosaic type), copy number variations (CNVs), single nucleotide variants (SNVs) and insertions-deletions (INDELs). Genomic variants detected in 11 cases could explain the short femur phenotype.The overall diagnostic rate was 11/27, with pathogenic/likely pathogenic variants explaining 9 cases and variants of uncertain significance (VUS) accounting for 1 cases. In addition Chromosomal karyotype analysis revealed chromosome mosaicism explaining 1 case. The diagnostic rate of CNV detection (CMA/CNV-seq) was 2/27, while WES achieved a diagnostic rate of 10/27, with an incremental yield of 8/27 in cases with negative or inconclusive CNV results. Pathogenic mutations involved genes such as FGFR3, SHOX, DUOX2, CLCN5, and HBA1/HBA2. ConclusionTrio-WES significantly improves the genetic diagnostic rate for fetal short femur length, particularly in cases where chromosomal analysis fails to identify a cause. Karyotyping remains valuable for detecting low-level mosaicism in large-scale chromosomal abnormalities, thereby complementing WES and CNV detection. A stepwise or parallel genetic testing strategy is recommended in prenatal diagnosis to optimize diagnostic efficiency and support clinical decision-making.
4.Serum anti-Müllerian hormone levels among female undergoing premarital and preconception examinations
LI Wei ; WAN Huiqing ; HAN Haiyan ; YU Jiqi ; HUANG Yizhou ; WANG Feixue ; ZHOU Jianhong
Journal of Preventive Medicine 2026;38(2):187-190
Objective:
To analyze serum anti-Müllerian hormone (AMH) level and its influencing factors among female undergoing premarital and preconception examinations, so as to provide the evidence for optimizing reproductive health management measures.
Methods:
Female who received free premarital and preconception examinations and voluntarily underwent serum AMH testing at Women and Children's Health Center of Gongshu District, Hangzhou City from January 2022 to April 2025 were selected as study subjects. Demographic information and reproductive history were collected through questionnaire survey. Venous blood was collected to detect thyroid-stimulating hormone (TSH), alanine aminotransferase, hemoglobin (Hb), and AMH levels. The original AMH values did not follow a normal distribution, but followed a normal distribution after natural logarithm transformation, recorded as ln (AMH). Pearson correlation analysis was used to analyze the correlation between variables, and multiple linear regression model was used to analyze the influencing factors for ln (AMH) levels among female undergoing premarital and preconception examinations.
Results:
A total of 1 073 females undergoing premarital and preconception examinations were surveyed, with a mean age of (29.43±3.30) years. There were 959 females without pregnancy (89.38%) and 1 042 females without full-term live birth history (97.11%). The median AMH level was 3.85 (interquartile range, 3.63) ng/mL, and the mean ln (AMH) level was 1.27±0.75. The ln (AMH) levels were higher in females without pregnancy and without full-term live birth history (both P<0.05). The ln (AMH) level was negatively correlated with age (r=-0.260) and TSH (r=-0.093), and positively correlated with Hb (r=-0.260, -0.093, and 0.073, all P<0.05). Multiple linear regression analysis showed that age (β′=-0.253) and TSH (β′=-0.103) were statistically associated with ln (AMH) levels among female undergoing premarital and preconception examinations (both P<0.05). The associations of other variables with ln (AMH) levels were not statistically significant (all P>0.05).
Conclusions
Age and TSH level are influencing factors for AMH levels among female undergoing premarital and preconception examinations. AMH levels among female undergoing premarital and preconception examinations decrease with increasing age and increasing TSH level.
5.Research on the prevalence of overweight and obesity among children
Xinyi LIANG ; Jingnan CHEN ; Xuelian ZHOU ; Ruimin CHEN ; Jingsi LUO ; Rongxiu ZHENG ; Chunxiu GONG ; Chunlin WANG ; Zhe SU ; Mireguli MAIMAITI ; Yan LIANG ; Hui YAO ; Haiyan WEI ; Hongwei DU ; Shaoke CHEN ; Yu YANG ; Feihong LUO ; Pin LI ; Min ZHU ; Wei WU ; Ke HUANG ; Guanping DONG ; Junfen FU
Chinese Journal of Pediatrics 2025;63(6):612-619
Objective:To investigate the prevalence and risk factors of overweight and obesity among Chinese children aged 3-18 years from 11 provinces, antonomous regions, or municipalities.Methods:This national cross-sectional community health survey utilized a multistage stratified cluster-random sampling method to recruit 193 997 nationally representative participants from 11 provinces, autonomous regions, or municipalities between January 2017 and December 2019. All participants underwent physical examinations, and their caregivers completed questionnaires assessing participants′ dietary, lifestyle, familial, and perinatal information. Multilevel multinomial logistic regression models were employed to identify the potential risk factors.Results:The cohort comprised 193 997 children (102 178 boys, 91 819 girls),aged (10±4) years. Overall prevalence rates were 30 574(15.8%)overweight children and 17 217(8.9%) obesity children. Boys exhibited higher overweight and obesity rates than girls (17.0% (17 368/102 178) vs. 14.4% (13 206/102 178), 11.3% (11 553/91 819) vs. 6.2% (5 664/91 819), χ2=249.12,1 578.69,both P<0.001). The detection rates of obesity in Tanner stage 2 and 3 were the highest in boys and girls, with 13.4%(2 231/16 665) and 8.6%(880/10 221) respectively. Risk factors for obesity included parental overweight (paternal OR=2.34 and maternal OR=2.29), annual household income of 100 000-200 000 yuan (compared with<100 000 yuan, OR=1.04), higher paternal education (compared with below high school,high school and a college education OR=1.09,1.14), birth weight >4.0 kg (≤5 and>5 years old OR=1.74, 1.44,respectively), and western food consumption≥1 time/month (compared with<1, 1-2, 3-4,>4 times/month OR=1.36, 1.30, 1.67(≤5 years), 1.19, 1.16, 1.15 (>5 years), respectively) (all P<0.05). Conversely, coarse grain intake≥1 times/week (compared with<1 times/week, every day, 3-4, 1-2 times/week OR=0.74, 0.80, 0.71 (≤5 years), 0.75, 0.87, 0.90(>5 years), respectively, all P<0.05) was associated with reduced obesity risk. Conclusions:Obesity epidemiology in children demonstrates significant heterogeneity across age, gender, geographic regions, and pubertal stages. It is necessary to establish a personalized prevention and control strategy.
6.Clinical characteristics and efficacy of glyceryl phenylbutyrate treatment in 20 pediatric patients with urea cycle disorder
Wenjuan QIU ; Chengkai SUN ; Yuan XIAO ; Xiaoyan HUANG ; Cui SONG ; Jin WU ; Haiyan WEI ; Liwen WU ; Dan YU ; Hongwei DU ; Chen LIU ; Xuefan GU
Chinese Journal of Pediatrics 2025;63(9):1005-1010
Objective:To investigate the clinical characteristics of urea cycle disorder (UCD), the efficacy and safety of glyceryl phenylbutyrate (GPB) therapy in pediatric patients with UCD.Methods:This study was a retrospective, single-arm, multicenter clinical study. The clinical data of 20 pediatric patients with UCD who received GPB treatment at 9 hospitals nationwide between December 2021 and August 2024 were collected. The clinical manifestations, laboratory results, and molecular genetic characteristics were analyzed, ammonia levels and other laboratory results were evaluated pre-post GPB therapy by paired t-tests or Wilcoxon tests. Results:Among the 20 pediatric patients with UCD, there were 8 males and 12 females, and the onset age was 2.8 (1.4, 5.7) years. The ammonia levels were 174 (125, 342) μmol/L at first onset. The symptoms included vomiting in 6 cases, drowsiness in 5 cases, epilepsy in 5 cases, developmental delay in 5 cases, psychiatric and behavioral abnormalities in 3 cases, and lethargy in 1 case, and 18 cases exhibited abnormal liver function. Twenty cases included 6 UCD subtypes, with 11 cases being ornithine transcarbamylase deficiency. A total of 27 variants were identified, 11 (41%) of which were novel. The age of patients who began GPB therapy was 4.0 (1.5, 6.6) years. Ten cases stopped GPB after 4.2 (3.4, 5.3) months, with 4 patients undergoing liver transplantation and 6 discontinuing for financial reasons. The remaining ten patients continued GPB therapy for 11.6 (8.6, 14.0) months. The duration of GPB treatment was 6.0 (4.2, 12.3) months, at the final visit, the levels of ammonia, platelets and aspartate aminotransferase were lower compared to those of pre-treatment (all P<0.05). The serum albumin level was higher than that of pre-treatment ( P=0.016). Two patients suffered only one episode of acute hyperammonaemia, with ammonia levels of 232 and 141 μmol/L, respectively. Nine cases experienced adverse effects potentially related to GPB, decreased appetite in 6 cases, vomiting in 3 cases, abnormal skin oil odor in 2 cases, somnolence, fatigue and diarrhea each in 1 case, with symptoms improved within 6 (3, 10) days. Conclusions:UCD primarily manifests with neurological and gastrointestinal symptoms, and early diagnosis of UCD could be achieved through the analysis of ammonia. GPB may effectively reduce ammonia levels in UCD pediatric patients, with favorable safety and tolerability.
7.Early differentiation of Kawasaki disease shock syndrome and septic shock in children
Haiyan GE ; Shuang LIU ; Jing CHEN ; Wenping GAO ; Siyuan HUANG ; Fang LI ; Fang LYU ; Dong QU
Chinese Journal of Pediatrics 2025;63(11):1229-1233
Objective:To explore the differences in early clinical features between Kawasaki disease shock syndrome (KDSS) and septic shock (SS).Methods:A retrospective case-control study was conducted. Clinical data was collected from 64 children who were diagnosed with KDSS or SS and admitted to the Department of Critical Care Medicine of Capital Center for Children′s Health, Capital Medical University from January 2018 to February 2025. Mann-Whitney U test, χ2 test, or Fisher′s exact test were used to compare the differences in clinical features, treatment, and outcomes between children with KDSS and SS. Lasso regression was applied to screen predictive variables, and multivariable logistic regression analysis was performed to identify factors associated with KDSS. Receiver operating characteristic (ROC) curve was used to evaluate the predictive value of parameters for KDSS. Results:Among the 64 children (30 males and 34 females), the age was 3.6 (1.2, 6.5) years. There were 51 cases in the SS group and 13 cases in the KDSS group. Compared to children with SS, children with KDSS had a longer pre-shock fever duration, lower lactate levels and serum albumin levels, and higher soluble interleukin-2 receptor (sIL-2R) levels (all P<0.05). Additionally, they exhibited a higher incidence of coronary involvement, pericardial effusion, and ascites, a higher utilization rate of intravenous immunoglobulin, and a lower utilization rate of invasive mechanical ventilation (all P<0.05). There was no significant difference in in-hospital mortality between KDSS and SS ( P=0.574). Multivariate logistic regression analysis identified pre-shock fever duration and sIL-2R as independent factors associated with KDSS ( OR=1.52 and 1.54 per 1 000 U increase, 95% CI 1.12-2.05 and 1.06-2.24, respectively; both P<0.05). ROC curve analysis showed that the areas under the curve for pre-shock fever duration and sIL-2R in identifying KDSS were 0.83 (95% CI 0.73-0.94, P=0.001) and 0.70 (95% CI 0.53-0.87, P=0.042), respectively. The optimal cutoff values were 3.5 d and 3.8×10 6 U/L, with sensitivities of 0.91 and 0.82, and specificities of 0.71 and 0.62, respectively. Conclusions:Children with KDSS have higher incidences of coronary involvement, pericardial effusion, and ascites compared to those with SS. Pre-shock fever duration and sIL-2R may serve as potential early indicators for distinguishing KDSS from SS.
8.Research advances on chondrocyte apoptosis in osteoarthritis
Jiwei HUANG ; Longfei WU ; Yuhao ZHAO ; Haiyan ZHAO
Chinese Journal of Orthopaedics 2025;45(18):1217-1226
Osteoarthritis is a chronic inflammatory disease characterized by damage to the articular cartilage, synovitis, and subchondral bone remodeling. Its pathological mechanisms involve extracellular matrix degradation, cell apoptosis, autophagy, and inflammatory responses. Among these, dysregulated apoptosis is a central driver of disease progression, making chondrocyte apoptosis a critical therapeutic target. This review summarizes current understanding of OA pathogenesis. Pro-inflammatory cytokines [e.g., interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and IL-6] exacerbate cartilage catabolism by activating signaling pathways like NF-κ B and MAPK. Chemokines, including the C-C motif chemokine ligand (CCL) family and the C-X-C motif chemokine ligand (CXC) family, amplify the inflammatory cascade by recruiting inflammatory cells, thereby contributing to the pathological process of osteoarthritis. In the study of programmed cell death, apoptosis is divided into extrinsic (death receptor pathway) and intrinsic (mitochondrial pathway) types. Both pathways induce chondrocyte apoptosis by activating the caspase cascade. Reactive oxygen species and inflammatory factors can promote excessive chondrocyte apoptosis through these pathways. Therapeutic strategies targeting apoptosis are diverse and include non-coding RNAs (miRNA, lncRNA, circRNA) that inhibit apoptosis by regulating related signaling pathways; phytochemicals that exert anti-inflammatory and anti-apoptotic effects; exosomes that suppress apoptosis by modulating immune responses and metabolism; and proteins/cytokines as well as melatonin, which protect chondrocytes by regulating specific signaling pathways. Clinical studies suggest these approaches hold promise for precision and personalized therapy, though challenges such as high cost, off-target effects, and drug resistance remain. In addition, drug delivery systems based on biomaterials (hydrogels) and nanotechnology can improve drug bioavailability and targeting. For example, drug-loaded hydrogels enable sustained release, and nanoparticles enhance drug stability and delivery efficiency, offering new perspectives for the treatment of osteoarthritis.
9.Changes in the epidemiology and clinical characteristics of human metapneumovirus in children with acute lower respiratory tract infection following the withdrawal of non-pharmaceutical interventions
Ke HUANG ; Tingting LAN ; Nuo XU ; Ting ZHAO ; Haiyan LI ; Lin DONG
Chinese Journal of Infectious Diseases 2025;43(3):143-150
Objective:To investigate the changes of the epidemiology and clinical characteristics of human metapneumovirus (hMPV) among children with acute lower respiratory tract infection (ALRTI) before and after the discontinuation of non-pharmaceutical interventions (NPI) during coronavirus disease 2019 epidemic.Methods:This was a retrospective cohort study. Children hospitalized at The Second Affiliated Hospital and Yuying Children′s Hospital of Wenzhou Medical University between January 2021 and December 2023, who were diagnosed with ALRTI by nasopharyngeal secretion testing for respiratory pathogens nucleic acid were enrolled. Clinical and laboratory data were collected. Children admitted between January 1, 2021 and January 7, 2023 were classified as the pre-NPI withdrawal group (abbreviated as pre-withdrawal group), while those admitted from January 8, 2023 afterward were classified as the post-NPI withdrawal group (abbreviated as post-withdrawal group). Nasopharyngeal secretions from the enrolled children were tested for 13 respiratory pathogens using polymerase chain reaction-capillary electrophoresis fragment analysis, and bacterial cultures were also performed. Statistical analyses were performed using the Mann-Whitney U test or chi-square test. Results:A total of 30 855 ALRTI cases were enrolled, with 1 679 of hMPV-positive. In the pre-withdrawal group, there were 861 cases with an age of onset of 2.0(1.0, 3.0) years, and the highest proportion was in the 1 to <3 years age group, accounting for 35.3%(304/861). In the post-withdrawal group, there were 818 cases with an age of onset of 3.0(2.0, 4.0) years, and the highest proportion was in the 3 to <5 years age group, accounting for 39.2%(321/818).The age of onset in the post-withdrawal group was significantly older than that in the pre-withdrawal group ( Z=7.69, P<0.001) .The hMPV detection rate was higher in the pre-withdrawal group than that in post-withdrawal group (5.75%(861/14 984) vs 5.15%(818/15 871); χ2=5.25, P=0.022). In the pre-withdrawal group, the epidemic peaks occurred in winter and spring, with the highest rates in January 2022(25.2%(224/890)) and March 2022 (21.6%(186/860)). In the post-withdrawal group, the epidemic peak shifted to spring and summer, and the detection rate became increased since April 2023(10.8%(136/1 258)). The post-withdrawal group showed lower rates of wheezing, shortness of breath, cyanosis, respiratory support, severe pneumonia, intensive care unit admission, and shorter hospital stays compared to the pre-withdrawal group ( χ2=69.09, 31.63, 12.97, 57.96, 55.73, 5.48 and Z=7.11, respectively, all P< 0.05).In the pre-withdrawal group, 412 cases (47.9%(412/861)) had other pathogens detected, compared to 445 cases (54.4%(445/181)) in the post-withdrawal group, indicating a significantly higher rate of co-infections in the post-withdrawal group ( χ2=7.20, P<0.05). The most commonly detected pathogens in both groups were Mycoplasma pneumoniae (MP), rhinovirus, and Streptococcus pneumoniae. However, the post-withdrawal group showed significantly higher detection rates of MP and influenza virus, but lower bacterial detection rates compared to the pre-withdrawal group ( χ2=39.41, 9.70, 5.63, respectively, all P<0.05). The detection rate of Haemophilus influenzae was 2.1%(17/818) in the post-withdrawal group which lower than that (6.7%(58/861)) in the pre-withdrawal group, and the difference was statistically significant ( χ2=21.32, P<0.001). Conclusions:In 2023, following the withdrawal of NPI, the epidemic peak of hMPV in Wenzhou area is delayed to spring and summer. The age of children with hMPV-associated ALRTI increases, with the majority being 3 to <5 years old. The overall severity of the disease decreases. However, the detection of mixed pathogens increases, with MP being the most common, while bacterial detection decreases.
10.Exploration on the Mechanism of Sanzi Sijun Formula in Non-alcoholic Fatty Liver Disease Based on Network Pharmacology and Experimental Validation
Junyao DING ; Ping HUANG ; Tao LIU ; Lili YANG ; Haiyan SONG ; Peiyong ZHENG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(11):30-39
Objective To explore the effects and mechanisms of Sanzi Sijun Formula(SSF)in non-alcoholic fatty liver disease(NAFLD)through network pharmacology,molecular docking and molecular dynamics simulation;To carry out experimental validation in vivo and in vitro.Methods The active components and target genes of SSF were screened using TCMSP,TCMIP and TCMIO databases.NAFLD-related targets were screened using the GeneCards database,and the intersection targets were obtained to construct a protein-protein interaction network and screen for core targets.The intersection targets were imported into the DAVID database for GO and KEGG enrichment analysis.Molecular docking was performed using AutoDock Vina software between the key active components of SSF and core targets,and molecular dynamics simulations were conducted using Gromacs 2022 for 100 ns.C57BL/6J mice NAFLD model was established by diet induction.SSF was administered by gavage for 8 weeks.Liver histopathological changes and the levels of non-esterified fatty acids(NEFA)were detected.In vitro NAFLD model was established by inducing AML12 cells with palmitic acid(PA)for 24 hours.SSF-containing serum was added to incubate simultaneously.The lipid accumulation and cell viability were detected.The core targets of SSF intervention in the in vitro and in vivo NAFLD models were verified by RT-qPCR and Western blot.Results Network pharmacological analysis identified 75 active components in SSF and revealed 179 shared targets between these components and NAFLD.Ten main active components including arachidonate,12-senecioyl-2E,8E,10E-atractylodin,cerebrosterol,glycyrrhizol B and sinapic acid,etc.as well as 8 core targets were identified.GO enrichment analysis of targets mainly involved protein phosphorylation,inflammatory response,and apoptosis,while the KEGG enrichment analysis mainly included AGE-RAGE,TNF,AMPK,PPAR and NF-κB signaling pathways.Molecular docking demonstrated that the major active components of SSF exhibited favorable binding affinity and stability with the core targets.Molecular dynamics simulation confirmed the stability of the complex of glyasperin B with AKT1,SIRT1,STAT3,PPARG,and TNF.SSF alleviated the pathological damage of liver tissues in mice NAFLD model,reduced NAS score and NEFA levels in liver tissues(P<0.05).Additionally,SSF reversed lipid accumulation and decreased cell viability of PA-induced AML12 cells(P<0.01).Further in vivo and in vitro experiments demonstrated that SSF significantly reversed the elevated mRNA levels of TNF-α,IL-6,IL-1β and PPARγ and protein expression of STAT3(P<0.05,P<0.01)in NAFLD models,up-regulated the protein levels of SIRT1 and p-Akt/Akt(P<0.05,P<0.01).Conclusion SSF can improve NAFLD of both in vitro and in vivo models.The regulation of multiple targets,such as AKT,SIRT1,STAT3 and PPARG,by its multiple active components,and adjustment of multiple approaches,such as lipid metabolism disorder,inflammatory responses,are involved in the potential underlying mechanisms.


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