1.Mechanism of Qingrun Prescription-containing Serum Improving Insulin Resistance in HepG2 Cells via Branched-chain α-keto Acid Dehydrogenase Regulation of Branched-chain Amino Acids (BCAAs)/mTOR Pathway
Xiangwei BU ; Xiaohui HAO ; Runyun ZHANG ; Meizhen ZHANG ; Ze WANG ; Haoshuo WANG ; Jie WANG ; Qing NI ; Lan LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):90-98
ObjectiveTo investigate the effect of Qingrun prescription(QRP)-containing serum on improving insulin resistance in HepG2 cells and its potential mechanisms. MethodsAn insulin resistance model was established in HepG2 cells with 1×10-6 mol·L-1 insulin. Branched-chain α-keto acid dehydrogenase (BCKDH) gene silencing was achieved using siRNA, and the cells were divided into 8 groups: normal group, model group (1×10-6 mol·L-1 insulin), metformin group (1 mmol·L-1 metformin), high-, medium-, and low-dose QRP groups (20%, 10%, and 5% QRP-containing serum, respectively), QRP + siRNA-silenced BCKDH (si-BCKDH) group (10% QRP-containing serum + si-BCKDH), and QRP + si-NC group (10% QRP-containing serum + si-NC). Glucose levels in the supernatant were measured with a glucose assay kit, while glycogen content was assessed using a glycogen assay kit. Levels of branched-chain amino acids (BCAAs) and branched-chain keto acids (BCKAs) were determined using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). mRNA transcription and protein expression levels of BCKDH, dishevelled, Egl-10, and pleckstrin (DEP) domain-containing mammalian target of rapamycin (mTOR)-interacting protein (DEPTOR), mTOR, and ribosomal protein S6 kinase 1 (S6K1) were detected using real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot. ResultsCompared to the normal group, the model group exhibited significantly decreased glucose consumption and glycogen content, increased levels of BCAAs and BCKAs, downregulated expression of BCKDH and DEPTOR, and upregulated mTOR and S6K1 expression (P<0.01). In comparison to the model group, QRP treatment at all doses significantly enhanced glucose consumption and glycogen content while reducing BCAAs and BCKAs levels (P<0.01). The high- and medium-dose QRP groups demonstrated significant upregulation of BCKDH mRNA transcription and protein expression, as well as DEPTOR mRNA transcription. Moreover, the DEPTOR protein expression level was significantly increased in high-, medium-, and low-dose QRP groups, while mTOR and S6K1 mRNA and protein expression levels were markedly downregulated (P<0.05, P<0.01). Compared to the QRP + si-NC group, the QRP + si-BCKDH group exhibited increased BCAAs and BCKAs levels, significantly decreased BCKDH mRNA transcription and protein expression, downregulated DEPTOR mRNA and protein expression, and upregulated mTOR and S6K1 mRNA and protein expression (P<0.05, P<0.01). ConclusionQRP may improve insulin resistance by reprogramming BCAAs metabolism. This effect involves upregulating BCKDH, reducing BCAAs and BCKAs levels, and suppressing the mTOR pathway activation.
2.Reliability and validity of general procrastination scale in the application of middle school students
Yongmei WU ; Yu CHEN ; Yunjia XIE ; Jili ZHANG ; Tianyi BU ; Jiawei ZHOU ; Zhengxue QIAO ; Jiarun YANG ; Xiaohui QIU ; Yanjie YANG
Chinese Journal of Behavioral Medicine and Brain Science 2024;33(2):161-165
Objective:To test the reliability and validity of the general procrastination scale (GPS) in the application of middle school students.Methods:The Chinese version of GPS, the irrational procrastination scale(IPS), and the Maslach burnout inventory(MBI) were utilized to survey 10 825 middle school students in Harbin City through stratified random sampling, and 4 498 students were retested after 4 weeks. Statistical analysis was performed using SPSS 27.0 and Mplus 8.0.Results:The entries were well differentiated.Exploratory and confirmatory factor analysis indicated that GPS was composed of two factors, including active avoidance and lack of planning.The model fit was good (CFI=0.914, TLI=0.901, RMSEA=0.069, SRMR=0.072). GPS was positively correlated with the total scores of IPS and MBI ( r=0.753, 0.677, both P<0.001). The Cronbach's α coefficient of GPS was 0.864, the folded half reliability was 0.870, and the retest reliability after 4 weeks was 0.756. Conclusion:The GPS has good reliability and validity among middle school students, which provides a standard for measuring the procrastination level of middle school students and carrying out related research.
3.Effects of microglia in the pathogenesis of major depressive disorder
Yanjie YANG ; Jili ZHANG ; Tianyi BU ; Kexin QIAO ; Xiaohui QIU ; Zhengxue QIAO ; Yu WANG ; Yu CHEN ; Bowen WAN ; Zihang XU
Chinese Journal of Behavioral Medicine and Brain Science 2024;33(3):278-283
Major depressive disorder (MDD) has become an increasingly serious public health issue, characterized by high incidence and high disability rates. It often coexists with other mental health problems and physical diseases, with a significant negative impact on patients' quality of life. In clinical practice, MDD is considered a heterogeneous disease. The complexity of the pathological mechanisms and the variability in treatment responses lead to a lack of clear therapeutic targets, which complicates the treatment process. In recent years, with advancements in neuroscience, the crucial role of microglia in the pathogenesis of MDD has been revealed. As the main immune cells in the brain, microglia are not only involved in the regulation of neuroinflammation but also play important roles in neurogenesis and neuronal regulation in MDD. This article mainly discusses the role of microglia in the pathophysiological mechanisms of MDD, aiming to provide a theoretical basis for microglia as a potential target for the treatment of MDD.
4.Effect of parenting style on social anxiety of college students: the mediating roles of core self-evaluation
Yu LIU ; Shaomin ZHANG ; Yanjie YANG ; Zhengxue QIAO ; Jiawei ZHOU ; Xiaomeng HU ; Tianyi BU ; Xuan LIU ; Kexin QIAO ; Xiaohui QIU
Chinese Journal of Behavioral Medicine and Brain Science 2024;33(7):642-646
Objective:To explore the impact of parenting style on social anxiety among college students, and examine the mediating effect of core self-evaluation.Methods:From November 2022 to January 2023, a total of 1 126 college students in Harbin were taken as research subjects.Interaction anxiousness scale(IAS), short-egna minnen betraffende upfostran-Chinese(s-EMBU-C) and core self-evaluations scale(CSES) were used for analysis. Data were analyzed using SPSS 26.0 software for correlation analysis and analysis of variance.AMOS 27.0 software was used for mediation effect test.Results:Social anxiety (42.31±8.23) was negatively correlated with positive parenting style (5.44±1.45) ( r=-0.072, P<0.05) and core self-evaluation (32.12±6.01) ( r=-0.350, P<0.01), while positively correlated with negative parenting style (7.40±1.74)( r=0.302, P<0.01). Core self-evaluation was positively correlated with positive parenting style ( r=0.362, P<0.01) and negatively correlated with negative parenting style ( r=-0.346, P<0.01).Parent parenting styles had a significant mean direct effect on social anxiety of college students ( βpositive=0.098, βnegtive=0.222).Mediation analyses indicated that core self-evaluation played a masking role between positive parenting styles and social anxiety, with an absolute value of 90.82% for the ratio of indirect(-0.089) to direct effects(0.098).Core self-evaluation had partial mediating effect on negative parenting styles, with direct effect and indirect effect accounting for 73.03% and 26.97% of the total effect, respectively. Conclusion:Parenting style can either directly affect college students' social anxiety or indirectly through the mediating effect of core self-evaluations, with core self-evaluations playing a masking role in the positive parenting styles pathway.
5.Bioinformatics analysis of regulatory network of long non-coding RNA LOC107987438 in depressive disorder
Tianyi BU ; Kexin QIAO ; Yan WANG ; Jili ZHANG ; Xiaohui QIU ; Zhengxue QIAO ; Jiawei ZHOU ; Jiarun YANG ; Wenjuan HE ; Yanjie YANG
Chinese Journal of Behavioral Medicine and Brain Science 2023;32(8):714-720
Objective:To investigate the regulatory role of defferentially expressed LOC107987438 in the pathogenesis of depressive disorder and provide a theoretical basis for its clinical application in depressive disorder.Methods:Differential expression of LOC107987438 was verified by quantitative real-time polymerase chain reaction(qRT-PCR)in peripheral blood monocular cells(PBMCs)of 60 patients with depressive disorder and 60 health controls. In addition, its diagnostic value was assessed by receiver operating characteristic(ROC)curves. Based on the ceRNA mechanism of lncRNA, the miRDB database was applied to predict the target miRNAs of LOC107987438, and the miRNAs with target score ≥ 80 among them were screened out.The screened miRNAs were then used to predict their potential target mRNAs through four databases which were TargetScan 8.0, miRTarBase, mirDIP and miRPathDB. Moreover, the predicted target mRNAs were annotated for gene ontology(GO)function annotation and tokoyo encyclopedia of genes and genomes(KEGG) pathway enrichment analysis via ClusterProfiler 4.0.5 package of R 4.1.1. Finally, a protein-protein interaction network was constructed using the STRING 11.5 platform to retrieve the interacting genes.Results:The qRT-PCR results showed that normalized expression of LOC107987438 in PBMCs of patients with depressive disorder was higher than that in health controls(depressive disorder: 2.084±1.357, health controls: 1.000±0.660, P<0.001). The ROC curve results showed that the area under curves(AUC)of LOC107987438 was 0.759(95% CI: 0.675-0.842, P<0.05), indicating its high potential diagnostic value. Bioinformatics analysis showed that hsa-miR-4670-3p, hsa-miR-619-3p, hsa-miR-6721-5p and hsa-miR-297 were the miRNAs with high bindings to LOC107987438. The results of KEGG signaling pathway enrichment revealed that hypoxia-inducible factor 1(HIF-1)signaling pathway, phosphatidylinositol 3-kinase-AKT(PI3K-Akt) signaling pathway and erythroblastic oncogene B(ErbB) signaling pathway were closely associated with depressive disorder. Among the top ten key genes screened by the protein-protein interaction network, kirsten rats arcomaviral oncogene homolog(KRAS), androgen receptor(AR), cyclic-AMP response binding protein1(CREB1), insulin-like growth factor 1(IGF1), cyclin-dependent kinase inhibitor 1B(CDKN1B) and calcium/calmodulin-dependent protein kinase type Ⅱ alpha(CAMK2A)were strongly associated with depressive disorder. Conclusion:The establishment of ceRNA regulatory network of LOC107987438 provides a theoretical basis for exploring the pathophysiology of depressive disorders.
6.Effect of ergosterol peroxide derivatives on the proliferation ,migration and invasion of human triple negative breast cancer cell
Hongyu ZHANG ; Wenkang REN ; Yu ZOU ; Yinglong HAN ; Hongyan YANG ; Ming BU ; Xiaohui DU ; Yu LIN
China Pharmacy 2022;33(11):1355-1360
OBJECT IVE To study the effects of ergosterol peroxide derivatives EP-3P on the proliferation ,migration and invasion of human tripe negative breast cancer cell MDA-MB- 231,and to provide reference for the development of breast cancer related drugs. METHODS MTT assay was adopted to detect the proliferation of MDA-MB- 231 cells after treated with 0(blank control),1.25,2.5,5,10,20,40 μmol/L EP-3P for 24,48 and 72 h. Wound healing assay and Transwell chamber method were adopted to detect the migration and invasion ability of MDA-MB- 231 cells after treated with 0(blank control ),5,10,20 EP-3P for 24 h. The apoptosis and cell cycle distribution were detected by flow cytometry. Western blot assay was used to detect the expressions of B-cell lympho ma-2(Bcl-2),Bcl-2 associated X protein (Bax),caspase-3,cleaved-caspase-3,cytochrome C (Cyt-C),matrix metalloproteinase- 2(MMP-2)and MMP- 9. RESULTS Compared with blank control group ,2.5,5,10,20,40 μmol/L EP-3P could significantly increase the inhibitory rate of cell proliferation (P<0.05 or P<0.01)in a dose and time- dependent manner. After 24 h treatment of EP- 3P(10,20 μmol/L),the rate of cell migration and the number of invasive cells were decreased significantly (P<0.01),and cell was arrested at G 2/M stage (P<0.05 or P<0.01);the apoptotic rate was increased significantly (P<0.05);the protein expressions of Bax ,Cyt-C and cleaved-caspase- 3 were upregulated significantly , while those of Bcl- 2,caspase-3,MMP-2 and MMP- 9 were downregulated significantly (P<0.01). CONCLUSIONS EP-3P can inhibit the proliferation ,migration and invasion of human tripe negative breast cancer cells MDA-MB- 231 through mitochondrial mediated endogenous caspase pathway ,and induce the apoptosis of cells .
7.Clinical observation on repairing nasal defect with multiple flaps
Ming DU ; Xia YANG ; Mingqiang BU ; Xiaohui DANG
Chinese Journal of Postgraduates of Medicine 2022;45(11):1038-1042
Objective:To summarize the clinical application of multiple flaps in repairing nasal defects.Methods:The clinical data of 32 nasal defect patients underwent flap repair in the Tengzhou Central People′s Hospital from 2017 to 2021 were analyzed retrospectively.Results:The nasal median displacement flap was used in 9 cases, nasofacial groove displacement flap in 10 cases, nasolabial groove flap in 8 cases, nasolabial displacement flap in 2 cases, frontal displacement flap in 2 cases, and free tragus cartilage flap in 1 case. After operation, all flaps survived, blood supply was good, nasal shape and stereoscopic structure returned to normal. There was no tumor recurrence during 6 to 24 months after operation.Conclusions:It is a good choice to use multiple flaps to repair the small defects of nasal parts. And the operation is simple and easy with high success rate.
8.Bioinformatics analysis of peripheral blood circRNAs in female patients with major depressive disorder
Wenxin ZHANG ; Yunmiao YU ; Tianyi BU ; Xiaohui QIU ; Xiuxian YANG ; Zhengxue QIAO ; Mingzhe ZHAO ; Yanjie YANG
Chinese Journal of Behavioral Medicine and Brain Science 2020;29(8):725-729
Objective:To screen the circRNAs with differential expression between female patients with major depressive disorder and healthy women, and to explore the circRNAs that might be associated with depressive disorder through bioinformatics analysis.Methods:Using high-throughput sequencing screen differentially expressed circRNAs of female major depressive disorder patients, |log 2FC|≥1 and FDR<0.05 were used to determine whether circRNA had a difference in expression.According to the miRNA sponges function of circRNA, the online databases were used to predict miRNAs which may be targeted by circRNAs, and miRNAs related target genes were also predicted by the five most different circRNA.GO and KEGG pathway enrichment analysis were used to predict biological processes for the target genes and signaling pathways.Based on the results of high-throughput sequencing, biological processes and signaling pathways related to depression, circRNAs related to depression were screened. Results:Thirteen targeted miRNAs were predicted by the five most different circRNAs(hsa_circ_0020959, hsa_circ_0005959, hsa_circ_0033064, hsa_circ_0006862, hsa_circ_0027732), and multiple biological processes and signaling pathways related to depressive disorders were predicted by the target genes, such as glucocorticoid receptor signaling pathway, interleukin-7 response, nervous system development, Wnt signaling pathway, FoxO signaling pathway, thyroid hormone signaling pathway, neurotrophin signaling pathway, and so on.Conclusion:All the 5 circRNAs enriched biological processes or signaling pathways related to depressive disorder, among which hsa_circ_0005959, hsa_circ_0033064, hsa_circ_0006862 and hsa_circ_0027732 may be more closely related to female major depressive disorder.
9.Network correlation of circRNA-miRNA and regulatory mechanisms of female major depressive disorder
Yunmiao YU ; Tianyi BU ; Xiaohui QIU ; Xiuxian YANG ; Zhengxue QIAO ; Erying ZHAO ; Jiawei ZHOU ; Wenxin ZHANG ; Ye SUN ; Yanjie YANG
Chinese Journal of Behavioral Medicine and Brain Science 2020;29(11):1007-1013
Objective:To obtain differential expression profiles of circRNAs and miRNAs in peripheral blood of female patients with major depressive disorder based on high-throughput sequencing technology, and then construct an interaction network.Based on the outcome, it made a further exploration of the possible occurrence and development mechanisms of female major depressive disorder through functional annotation and pathway enrichment analysis.Methods:According to the ceRNA theory, circRNA-miRNA network was constructed by TargetScan software via predicting the binding sites.Subsequently, the GO functional enrichment analysis was performed, and KEGG pathway enrichment analysis were utlized to illustrate the target genes of co-expressed miRNAs.Thereby, the key genes related to major depressive disorder could be screened out.Results:A total of 724 differential circRNAs and 26 differential miRNAs were detected in female patients with major depressive disorder.And hsa_circ_0086092 and hsa-miR-146a-3p were the most co-expressed.Go functional annotations pointed out that it involved the regulation of nucleobase-containing compound metabolic process, regulation of RNA splicing, regulation of cell communication, amino acid transfer, regulation of RNA metabolic process, regulation of signaling and other biological processes.KEGG pathway analysis showed that target genes were mainly enriched in neurotrophin signaling pathway, Rap1 signaling pathway, FoxO signaling pathway, AMPK signaling pathway, cocaine addiction, mTOR signaling pathway, Jak-STAT signaling pathway, cAMP signaling pathway, etcetera.Among the predicted target genes, BDNF, FGF2, MAPK14, GRIN2A, GRIN2B, GRM2 and PDE4 have the highest correlation with major depressive disorder.Conclusion:Hsa_circ_0086092 may be involved in the occurrence and development of female major depressive disorder through interaction with hsa-miR-146a-3p.
10. Aspirin intervenes in hyperlipidemia kidney damage by blocking endoplasmic reticulum stress in podocytes
Yudong CHU ; Rongshan LI ; Yuan TIAN ; Pengjie XU ; Jiang LIU ; Xiaohui QIU ; Shizhong BU
Chinese Journal of Nephrology 2020;36(2):139-144
Objective:
To investigate the effects and underlying mechanisms of aspirin on endoplasmic reticulum stress in podocytes induced by hyperlipemia.
Methods:
Cultured podocytes were divided into four groups: control group, aspirin (100 μg/ml) group, oxidized low density lipoprotein (ox-LDL, 100 μg/ml) group, aspirin+ox-LDL group. The expression of protein kinase R-1ike endoplasmic reticulum kinase (PERK), eukaryotic translation initiation factor 2α (eIF2α), activating transcription factor-4 (ATF4) and CAAT/enhancer binding protein homologous protein (CHOP) at 6 h, 12 h, 24 h, 48 h were evaluated by real-time PCR. The related proteins of p-PERK and p-eIF2α at 24 h and ATF4 at 12 h were evaluated by Western blotting, respectively.
Results:
The expressions of PERK, eIF2α peaked at 24 h, while ATF4 and CHOP peaked at 12 h in ox-LDL group and aspirin+ox-LDL group. Compared with control group, the expressions of PERK, eIF2α, ATF4 and CHOP were significantly higher in ox-LDL group at each times (all

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