1.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.
2.The Effects of Tai Chi Training on Bone Density,Bone Turnover Markers,and Heart Rate Variability in High-Risk Osteoporosis Population
Jiaming LIN ; Chao LI ; Wei ZHAO ; Jun ZHOU ; Xiaoying CHEN ; Xiangyu XI ; Haijun HE ; Baohong MI ; Yuefeng CHEN ; Weiheng CHEN
Journal of Traditional Chinese Medicine 2025;66(15):1566-1571
ObjectiveTo explore the effects of the Tai Chi training on bone density, bone turnover markers, and heart rate variability for people with high-risk osteoporosis, and to provide evidence for the prevention of osteoporosis at early stage. MethodsSixty-six cases of people with high risk of osteoporosis were included, and they were divided into 33 cases each in the intervention group and the control group using the random number table method. The control group received osteoporosis health education three times a week, and the intervention group received Tai Chi training under the guidance of a trainer three times a week for 40 mins each time on the basis of the control group, and both groups were intervened for 12 weeks. Dual-energy X-ray absorptiometry was used to measure the bone density of L1~L4 vertebrae, bilateral femoral necks and bilateral total hips in the two groups before and after the intervention; enzyme-linked immunosorbent assay was used to determine bone turnover markers before and after the intervention, including pro-collagen type Ⅰ pro-amino-terminal prepropyl peptide (P1NP) and β-collagen type Ⅰ cross-linking carboxy-terminal peptide (β-CTX). Seven cases with good compliance in the intervention group were selected. After wearing the heart rate sensor, they successively performed Tai Chi training and walking activities recommended by the guideline for 20 mins each, and the heart rate variability (HRV) during exercise was collected, including time-domain indexes such as standard deviation of normal sinus intervals (SDNN), root-mean-square of the difference between adjacent RR intervals (RMSSD), frequency-domain metrics such as low-frequency power (LF), high-frequency power (HF), and low-frequency/high-frequency power ratio (LF/HF), as well as nonlinear metrics such as approximate entropy (ApEn), sample entropy (SampEn). ResultsFinally, 63 cases were included in the outcome analysis, including 30 cases in the intervention group and 33 cases in the control group. After the intervention, the differences of L1~L4 vertebrae, bone density of bilateral femoral neck and bilateral total hip in the intervention group were not statistically significant when compared with those before intervention (P>0.05), while the bone density of all parts of the control group decreased significantly compared with that before intervention (P<0.05), and the difference in the bone density of the L1~L4 vertebrae, bilateral femoral neck, and the right total hip before and after the intervention of the intervention group was smaller than that of the control group (P<0.05). The differences in P1NP and β-CTX between groups before and after intervention was not statistically significant (P>0.05). Compared with walking exercise, LF decreased, HF increased and LF/HF decreased during Tai Chi exercise (P<0.05); the time domain indexes and non-linear indexes between groups had no statistically significant difference (P>0.05). ConclusionTai Chi exercise can maintain lumbar, hip, and femoral bone density and improve sympathetic/parasympathetic balance in people at high risk for osteoporosis, but cannot significantly improve bone turnover markers.
3.Effects of arsenic and its metabolites on p53 gene expression in BEAS-2B cells
Na LIU ; Jinjun JIANG ; He MA ; Ruihuan ZHAO ; Yuefeng HE ; Weihua WEN
Journal of Environmental and Occupational Medicine 2024;41(4):431-436
Background Arsenic is a human carcinogen. Arsenic and its metabolites affect the expression of p53, but whether there are any changes of p53 phosphorylation and ubiquitination levels in human bronchial epithelium cells (BEAS-2B) are not clear after exposure to arsenic and its metabolites. Objective To study the effects of arsenic and its metabolites monomethylarsic acid (MMA) and dimethylarsinic acid (DMA) on the expression of tumor suppressor gene p53 in BEAS-2B cells. Methods Different concentrations of sodium arsenite (NaAsO2) were used to infect BEAS-2B cells, and the cell viability was detected with CCK-8 reagent to determine the dose and time of NaAsO2 used for the following study. Based on the results of cell viability, the cells were divided into two panels: a sodium arsenide panel and an arsenic methylation metabolite penal. The doses of sodium arsenite were 0, 2, 4, and 6 μmol·L−1 NaAsO2; the arsenic methylation metabolite panel consisted of 0 μmol·L−1 NaAsO2 group (control), 6 μmol· L−1 MMA group, 6 μmol· L−1 DMA group, and 6 μmol· L−1 NaAsO2 group. The cells were collected after 48 h treatment, and the total protein and total RNA were extracted. The relative levels of p53 mRNA expression were determined by quantitative real-time polymerase chain reaction (qRT-PCR), the relative expression levels of p53 protein, p53 Ser9 and Ser15 phosphorylated proteins were determined by Western blot, and the level of p53 ubiquitination was detected by co-immunoprecipitation (CO-IP). Results Compared with the control group, the cell viability rates in all BEAS-2B cells treated by NaAsO2 were significantly reduced (P<0.05), and the 50% cell viability was observed at 6 μmol·L−1. Compared with the control group, the relative expression level of p53 mRNA gradually decreased after NaAsO2 (2, 4, 6 μmol·L−1) treatment (P<0.05), the relative expression levels of p53 protein and Ser9 phosphorylated protein induced by NaAsO2 also decreased gradually (P<0.05), and the relative expression level of p53 Ser15 phosphorylated protein induced by NaAsO2 followed the same pattern, but it was only lower than that of the control group in the 6 μmol·L−1 NaAsO2 group (P<0.05). Compared with the control group, there were no significant effects on the relative expression levels of p53 mRNA, p53 protein, Ser9 and Ser15 phosphorylated proteins in the MMA group and the DMA group. Compared with the control group, the expression level of p53 ubiquitination was significantly decreased and the expression of K48 ubiquitination decreased significantly after NaAsO2 infection. Conclusion Arsenic causes a decrease in the expression of the p53 protein in BEAS-2B cells, largely due to inhibition of the phosphorylated pathway and a decrease in mRNA expression, and protein changes caused by a decrease in p53 ubiquitination do not play a dominant role. MMA and DMA do not affect p53 gene expression.
4.Research on Material Basis of Homologous and Heterogeneous Effect of Aurantii Fructus Immaturus and Aurantii Fructus Based on Total Statistical Moment Analysis and Molecular Connectivity Index
Xiao YUAN ; Yuefeng WU ; Shixiong LI ; Youzhi LIU ; Zhiqin YU ; Fuyuan HE ; Jin ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(3):161-168
ObjectiveTo investigate the material basis of homologous and heterogeneous effect of Aurantii Fructus Immaturus(AFI) and Aurantii Fructus(AF) based on the total statistical moment analysis and molecular connectivity index(MCI). MethodRelevant literature at home and abroad and Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) were consulted to establish the chemical composition database of AFI and AF, and set up their fingerprints by ultra-high performance liquid chromatography(UPLC), and the total statistical moments and similarity parameters of the fingerprint were calculated. According to MCI, all components of AFI and AF were divided into different component groups, the average values of 0-8th order(0χ-8χ) MCI of the common component groups of AFI and AF were calculated. ResultThe values of total zero-order moment(AUCT) of AFI and AF were (10.57±2.45)×106, (5.09±0.89)×106 μV·s, the values of total first-order moment(MCRTT) were (11.57±1.58), (12.10±1.29) min, the values of total second-order moments(VCRTT) were(24.49±2.30), (26.49±2.54) min2, respectively. It showed that qualitative and quantitative parameters of AFI and AF were significantly different. The components with high similarity such as neohesperidin, hesperidin and narirutin were screened as the common potential pharmacodynamic components of AFI and AF. The non-common components of AFI, such as alysifolinone and imperatorin, and the non-common components of AF, such as neoeriocitrin and isosakuranin, with high similarity were screened out as potential heterogeneous components of AFI and AF. The composition groups of AFI and AF were classified into six categories, and the similarities between the composition groups of AFI and AF and the total constituents were 0.872-0.979 and 0.918-0.997, the average values of 0χ-8χ MCI of alkaloids in AFI and AF were 3.65 and 3.14, the average values of 0χ-8χ MCI of flavonoids were 8.47 and 8.47, the average values of 0χ-8χ MCI of volatile oils were 2.71 and 3.48, respectively. It showed that there were some differences in MCI of chemical constituents(groups) between AFI and AF. ConclusionThe chemical constituents(groups) of AFI and AF not only differ in content and species, but also in structural characteristics and structure-activity relationship, which can provide a basis for further explaining the scientific connotation of homologous and heterogeneous effect of AFI and AF.
5.Safety and efficacy of neoadjuvant chemotherapy combined with immunotherapy in 101 patients with muscle-invasive bladder cancer
Chaosheng GAN ; Tao LI ; Junjie FAN ; Zhangdong JIANG ; Guojing WANG ; Ke XU ; Qiyuan KANG ; Yangqingqing ZHOU ; Yuefeng DU ; Jinhai FAN ; Lei LI ; Dalin HE ; Kaijie WU
Journal of Modern Urology 2024;29(9):790-796
Objective To explore the safety and efficacy of neoadjuvant chemotherapy(NAC)combined with immunotherapy before radical cystectomy plus pelvic lymph nodes dissection(RC-PLND)for muscle-invasive bladder cancer(MIBC).Methods The clinical data of 101 patients with MIBC who underwent neoadjuvant therapy followed by RC-PLND in the Department of Urology,the First Affiliated Hospital of Xi'an Jiaotong University during Jan.2019 and Dec.2023 were retrospectively analyzed,including 71 patients(70.3%)who received NAC(NAC group)and 30(29.7%)who received NAC combined with immunotherapy(NAC combine immunotherapy group).The clinical and pathological data and adverse events during neoadjuvant therapy were compared.Logistic regression analysis was used to explore the independent predictors of pathological complete response(pCR)and pathological partial response(pPR).Results There were no significant differences in the baseline data between the two groups(P>0.05).However,the proportion of multiple tumors in patients receiving NAC before surgery was significantly higher than that in the NAC combined immunotherapy group(69.0%vs.46.7%,P=0.034).Compared with NAC group,NAC combined with immunotherapy group had significantly improved rate of pathological downstaging and pPR(60.6%vs.83.3%,P=0.026;45.1%vs.70.0%,P=0.022).Furthermore,the rate of pCR in patients undergoing NAC combined immunotherapy was higher than those undergoing NAC,but the difference was not significant(53.3%vs.33.8%,P=0.067).Logistic regression analysis revealed that clinical T-stage and tumor diameter were independent predictors of pCR and pPR(P<0.05).In addition,the most common adverse events during neoadjuvant therapy were anemia,decreased white blood cells,nausea,and vomiting,but most of them were grade 1-2 and could be relieved through symptomatic treatment.Conclusion NAC combined with immunotherapy is safe and effective,which can improve the rate of pathological downstaging,pPR and pCR,without increasing the incidence of adverse reactions.
6.Quality Assessment of Ginger and Its Processed Products Based on Fingerprints and Chemometrics
Xiujuan YANG ; Jianqiao HE ; Jingjing GUO ; Jiajia WANG ; Pengxian NIU ; Zhijun YANG ; Yuefeng LI ; Shou LI
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(10):1593-1599
Objective To establish the fingerprint map and method of multi-component content determination of ginger and its processed products,and to evaluate their quality. Methods High performance liquid chromatography (HPLC) was used to establish the fingerprint of ginger and its processed products (dried ginger,baked ginger,ginger charcoal) and the method for the simultaneous determination of content of five components. The data were analyzed by similarity evaluation and chemical pattern recognition. Results The fingerprint of ginger and its processed products was established. The similarity was ranged from 0.931 to 0.996. A total of 15 common peaks were confirmed. Five components (6-ginger phenol,8-ginger phenol,10-ginger phenol,6-ginger enol,zingiberone) were identified when compared with the standard. The content of 6-ginger phenol,8-ginger phenol and 10-ginger phenol in ginger and its processed products(dried ginger,baked ginger,ginger charcoal),which were determined by HPLC,met the standard of Chinese Pharmacopoeia(2020 edition). The content of 6-ginger enol and zingiberone in fresh ginger was very low,but their content increased significantly after processing into dried ginger and baked ginger. Ginger and its processed products can be divided into different category in cluster analysis. Five components with VIP greater than one were selected by orthogonal partial least squares discriminant analysis. Conclusion The HPLC fingerprint and assay method established in this study are specific,simple and feasible,stable and reliable. With the help of chemical pattern recognition method,it can provide reference for the quality evaluation of ginger and its processed products.
7.Chemical Constituents and Insulin Resistance Targets in Different Parts of Morus alba by UPLC-Q-TOF-MS Combined with Network Pharmacology
Yuefeng WU ; Jinling LIU ; Dingfang CHEN ; Zhifei ZHU ; Youzhi LIU ; Xiao YUAN ; Jin ZHOU ; Fuyuan HE
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(2):149-158
ObjectiveTo compare the similarities and differences of material basis for improving insulin resistance (IR) in different parts of Morus alba based on liquid-mass combination combined with network pharmacology and molecular docking technology. MethodUltra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was used to analyze the composition differences in different parts of M. alba. Sybyl-X2.1 was used to connect components with IR core targets, and the selection criterion was Total Score≥5. The "component-target-disease" network map was drawn. The total statistical moment standard similarity (TQSMSS) between the single target-component docking score data set and the total target-component docking score data set was calculated. The targets with higher TQSMSS were screened out, and the protein-protein interaction (PPI) network was constructed. The Gene Ontology (GO) functional analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed using R language. ResultForty-one active components were obtained by UPLC-Q-TOF-MS. According to the total statistical moment (similarity) method, there were 20, 23, 30, and 27 targets with TQSMSS≥0.75 in Mori Ramulus, Mori Cortex, Mori Fructus, and Mori Folium, respectively. In the four M. alba medicinal sources, the functional order of the targets by GO enrichment analysis was Mori Fructus>Mori Folium>Mori Cortex>Mori Ramulus, which were involved in biological processes such as blood glucose homeostasis, glucose metabolism, and glucose transmembrane transport. The order of the four M. alba medicinal sources by KEGG pathway enrichment analysis was Mori Fructus>Mori Ramulus>Mori Folium>Mori Cortex, which were involved in the adenosine monophosphate-activated protein kinase (AMPK) energy metabolism signaling pathway, the insulin regulation-related signaling pathway, the anti-inflammatory and anti-oxidative stress signaling pathway, and so on. ConclusionThis research demonstrates that there are differences in the material basis for improving IR by different parts of M. alba, which provides references for the development of different parts of M. alba.
8.Analysis on Quality Attribute of Substance Benchmarks of Shentong Zhuyutang Based on Supramolecular "Imprinting Template" Theory
Zhifei ZHU ; Youzhi LIU ; Yuefeng WU ; Xiao YUAN ; Jin ZHOU ; Fuyuan HE
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(11):171-178
ObjectiveBased on the supramolecular "imprinting template" theory, the autonomous action law of the component groups of Shentong Zhuyutang in the preparation process of medicinal materials-decoction pieces-formulas was studied to clarify the quantitative transfer law of its quality attributes. MethodUltra performance liquid chromatography(UPLC) fingerprint of Shentong Zhuyutang was established with mobile phase of 0.4% phosphoric acid aqueous solution(A)-acetonitrile(B) for gradient elution(0-2.5 min, 100%A; 2.5-6 min, 100%-96%A; 6-15 min, 96%-92%A; 15-25 min, 92%-88%A; 25-35 min, 88%-75%A; 35-50 min, 75%-65%A; 50-60 min, 65%-50%A; 60-65 min, 50%-30%A; 65-70 min, 100%A) and detection wavelength of 235 nm, and the total statistical moments, information entropy and primary feeding amount of fingerprint of medicinal materials, decoction pieces and benchmark samples were calculated. Dry extract rate of the benchmark samples, the transfer rates and the addition parameters of medicinal materials-decoction pieces-formulas were calculated. ResultSimilarities of the total statistical moments of UPLC fingerprint of 15 batches of medicinal materials and decoction pieces were>0.89, the relative standard deviations(RSDs) of information entropy of UPLC fingerprint of 12 medicinal materials and decoction pieces were<10%. RSDs of total first-order moment(MCRTT) and information entropy of Shentong Zhuyutang(medicinal materials) were 5.5% and 2.3%, while the RSDs of MCRTT and information entropy of Shentong Zhuyutang(decoction pieces) were 4.8% and 2.6%, respectively. The dry extract rate of 45 batches of Shentong Zhuyutang was 17.2%-20.2%. The transfer rate of medicinal materials to decoction pieces was within the range of data fluctuation, which was 70%-130% of the average value. The overall transfer rates of medicinal materials to decoction pieces and decoction pieces to benchmark samples were 101.8% and 83.0%, respectively. ConclusionThe quality properties of Shentong Zhuyutang benchmark samples can be studied by total statistical moment analysis and primary feeding amount analysis, which can confirm the supramolecular "imprinting template" theory to a certain extent.
9.Effects of arsenic and its metabolites on expressions of BCL-2α and BCL-2β transcripts
Xingyu YAN ; Zihan QU ; Huijie PU ; Xingquan YANG ; Tianlin ZHOU ; Yuefeng HE
Journal of Environmental and Occupational Medicine 2022;39(1):78-84
Background Arsenic is a toxicant that can affect the expressions of the cellular anti-apoptotic gene BCL-2 and its protein, but the effects of arsenic on BCL-2α and BCL-2
10.Discussion on Material Basis of Different Parts of Mulberry with Different Effects Based on Molecular Connectivity Index
Yuefeng WU ; Zhifei ZHU ; Jinling LIU ; Qimeng FAN ; Youzhi LIU ; Jin ZHOU ; Fuyuan HE
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(13):152-160
ObjectiveTo explore the material basis for the difference in the efficacy of different parts of mulberry based on molecular connectivity index (MCI). MethodBy referring to the relevant literature at home and abroad and traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) database, the chemical composition database of mulberry-source medicinal materials was established. Venn analysis was carried out on the components among mulberry-source medicinal materials. The components in the database were divided into 10 categories, and the composition information was analyzed. According to MCI value, all components of mulberry-source medicinal materials were divided into different groups. The angle cosine method was used to calculate the MCI similarity. The average MCI values of the common component group from 0-8 orders and CI of mulberry-source medicinal materials were calculated. ResultThe components with high similarity such as (+)-cycloolivil, 1′-methoxy-2′-hydroxydihydromollugin, kuwanon, morusin and 1-deoxynojirimycin were selected as potential pharmacodynamic components. Mulberry-source medicinal materials could be divided into five component groups. The similarity between component groups and total components was 0.760-0.999, and the similarity between component groups was 0.248-0.999. In Mori Ramulus, Mori Folium, Mori Cortex and Mori Fructus, the average MCI values of their flavonoids from 0-8 orders were 4.57, 4.59, 6.41, 4.24, respectively. The average MCI values of alkaloids from 0-8 orders were 2.65, 4.55, 2.58, 2.78, respectively. The average CI values from 0-8 orders were 5.51, 5.49, 5.44 and 2.88, respectively. ConclusionIt is preliminarily concluded that there are differences in the flavonoids and pathways of hypoglycemic effects between Mori Cortex and the other three mulberry-source medicinal materials. The MCI values of alkaloids from 0-8 orders in Mori Folium and Mori Fructus were higher, but their inhibitory activity of α-glucosidase were lower than those of Mori Ramulus and Mori Cortex. The structural characteristics of the total components of Mori Fructus represented by CI were quite different from the other three mulberry-source medicinal materials.

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