1.Methodological analysis of sequential sector scan through oral fissure for fetal cleft palate screening
Xiaofeng LU ; Yi ZHOU ; Wanyan LI ; Liang LI ; Chuanfen GAO ; Jingwen SHE ; Jing QIN ; Chaoxue ZHANG
Chinese Journal of Perinatal Medicine 2022;25(5):339-342
Objective:To explore the method of sequential sector scan with 2D ultrasound through oral fissure (SSTOF) and its utilization in the cleft palate screening.Methods:Based on features of oral anatomy and ultrasonic beam, SSTOF was designed to screen cleft palate and the accuracy had been verified using specimens of aborted fetuses. This study recruited 7 154 women with singleton pregnancy who were screened for fetal malformations during 20-28 gestational weeks in the First Affiliated Hospital of Anhui Medical University from May 2020 to October 2020. In medical addition to routine screening, these subjects also underwent SSTOF to further verify its feasibility and imaging performance. Follow-up was performed by telephone and medical record review.Results:Clear images of the upper palate were acquired in five specimens of induced fetuses using SSTOF. Except for 56 cases lost to follow-up, a total of 7 098 fetuses were finally enrolled, of which 6 885 acquired satisfactory images using SSTOF, 213 did not due to inappropriate position, with a success image rate of 97%. SSTOF found cleft palate in 31cases, which were all confirmed after birth or induction, noting an accuracy rate of 100%.Conclusion:Sequential sector scan through oral fissure has a high clinical value on cleft palate screening in the second trimester with advantages of clear image, easy operation, and access to section views.
2.Molecular Mechanism of Erxian Decoction in Treatment of Anxiety Disorder Based on Network Pharmacology and Experimental Verification
Kaijie SHE ; Jingwen YANG ; Danhua MENG ; Wenqing LIANG ; Zihan GONG ; Guangxin YUE
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(19):185-193
ObjectiveTo predict the potential molecular mechanism of Erxian decoction in the treatment of anxiety disorder based on network pharmacology, and to verify the efficacy and mechanism using the animal model of maternal separation combined with restraint stress. MethodActive components and related targets of Erxian decoction were obtained by traditional Chinese medicine system pharmacology database and analysis platform (TCMSP) and SwissTargetPrediction. The targets related to anxiety disorder were screened out through GeneCards, therapeutic target database (TTD), online mendelian inheritance in man database (OMIM), and DrugBank, and the drug-disease intersection targets were obtained by taking intersections with the drug targets. The protein-protein interaction (PPI) network was constructed by the STRING database, and the core targets were screened out based on topological parameter analysis. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were carried out for the intersection targets through the Metascape platform. Maternal separation combined with restraint stress was used to induce the mouse model of anxiety disorder. From the end of lactation on the 21st postnatal day (PD21) to the completion of restraint stress on the 97th postnatal day (PD97), the mice were fed with Erxian decoction mixed with diet. The anxiety state of mice was evaluated by open field test and elevated O-maze test. The content of plasma corticosterone (CORT) in mice was detected by enzyme-linked immunosorbent assay (ELISA). The expression levels of protein kinase B (Akt1), mammalian target of rapamycin (mTOR), brain-derived neurotrophic factor (BDNF), postsynaptic density-95 (PSD95), and synaptophysin in the hippocampus of mice were detected by Western blot and real-time quantitative polymerase chain reaction (Real-time PCR). ResultNinty-seven active components and 227 action targets of Erxian decoction were obtained. There were 3 863 targets related to anxiety disorder, with 161 drug-disease intersection targets. Among these intersection targets, core targets such as Akt1, interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor (TNF), and mTOR were presumedly closely related to anxiety disorder. The results of KEGG pathway analysis showed that Erxian decoction mainly treated anxiety disorder through phosphatidylinositol 3-kinase (PI3K)/Akt, mitogen-activated protein kinase (MAPK), and neuroactive ligand-receptor interaction signaling pathways. The results of animal experiments showed that compared with the model group, the Erxian decoction group significantly increased the time of mice spent in the central zone and central crossing times and time spent in the opened arm and opened arm crossing times, with significantly increased expression levels of p-Akt1, p-mTOR, BDNF, PSD95, and synaptophysin (Syp). ConclusionErxian decoction has the multi-target and multi-pathway characteristics in the treatment of anxiety disorder, and its mechanism may be related to the improvement of synaptic plasticity and neuroinflammation by affecting Akt1, IL-1β, IL-6, TNF, mTOR, and other core targets and modulating PI3K/Akt, MAPK, as well as neuroactive ligand-receptor interaction signal pathways.
3.Feasibility of Erxian Decoction and Wenshen Prescription in Treatment of Depression Based on Network Pharmacology and Experimental Verification
Kaijie SHE ; Zihan GONG ; Jingwen YANG ; Wenqing LIANG ; Danhua MENG ; Guangxin YUE
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(16):211-220
ObjectiveTo predict the molecular mechanism of Erxian decoction and Wenshen prescription (modified Erxian decoction) in the treatment of depression based on network pharmacology and explore the feasibility of Wenshen prescription in the treatment of depression by comparing the efficacy and mechanism of the two decoctions based on a depression model induced by maternal separation combined with chronic restraint stress. MethodActive components and targets of Erxian decoction and Wenshen prescription were collected through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Bioinformatics Analysis Tool for Molecular mechanism of Traditional Chinese Medicine (BATMAN-TCM). Targets related to depression were screened out from databases such as GeneCards, Online Mendelian Inheritance in Man database (OMIM), and DrugBank. Common targets of drugs and disease were obtained and imported to Cytoscape 3.8.2 to plot the drug-active component-target-disease network. STRING platform was used to construct a protein-protein interaction (PPI) network and core targets and related core components were screened out. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis were performed on common targets through Metascape platform. The depression model was induced in mice by maternal separation combined with chronic restraint stress. From the 21st day of maternal separation (PD21) to the 111th day of restraint stress completion (PD111), mice were fed with the diet mixed with Erxian decoction or Wenshen prescription for intervention. The depressive state of mice was evaluated according to the sucrose preference test, tail suspension test, open field test, and elevated O-maze test. The expression of ionized calcium-binding adapter molecule 1 (Iba1) in the microglia was observed by immunohistochemistry (IHC). Western blot and Real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR) were used to detect the expression levels of protein kinase B1(Akt1), brain-derived neurotrophic factor (BDNF), postsynaptic density-95 (PSD95), and synaptophysin (Syn). ResultA total of 126 and 118 targets of Erxian decoction and Wenshen prescription in the treatment of depression were screened out, with only eight more targets of Erxian decoction than Wenshen prescription. The two decoctions shared the same core targets, mainly including Akt1, interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α). KEGG pathway enrichment analysis predicted that Erxian decoction and Wenshen prescription mainly treated depression through the phosphatidylinositol-3 kinase (PI3K)/Akt signaling pathway, mitogen-activated protein kinase (MAPK) signaling pathway, and neuroactive ligand-receptor interaction pathway. Animal experiments showed that compared with the results in the model group, Erxian decoction and Wenshen prescription could up-regulate the sucrose preference index, prolong the time spent in the central zone, increase the number of crossings, prolong the time spent in opened arm, increase the number of crossings in the opened arm, elevate the expression levels of p-Akt1, BDNF, PSD95, and Syn (P<0.05, P<0.01), shorten the immobility time of tail suspension, and reduce the expression level of Iba-1 in the hippocampal microglia (P<0.05, P<0.01). No significant difference between the two decoctions was found. ConclusionUnder the pathogenesis and syndrome law of depression dominated by kidney yang deficiency, Wenshen prescription modified from Erxian decoction is feasible in the treatment of depression. The mechanism may be attributed to the fact that both decoctions can improve neuroinflammation and synaptic plasticity in the hippocampus by affecting Akt1, IL-1β, IL-6, TNF-α, and other core targets and regulating the PI3K/Akt, MAPK, and neuroactive ligand-receptor interaction signaling pathways.
4.Syndromes and Mechanisms of Depression Induced by Second Hit in Mice
Zihan GONG ; Ying WANG ; Jingwen YANG ; Wenqing LIANG ; Danhua MENG ; Kaijie SHE ; Yuan LIANG ; Guangxin YUE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(6):29-38
ObjectiveTo explore the syndromes and mechanisms of depression induced by maternal separation (MS) combined with chronic restraint stress (RS) in mice. MethodOn postnatal day 0 (PD0), the offspring mice were randomized into a blank group (NC) and a modeling group. The mouse model of depression was established by MS+RS for 21 days. After removal of female mice on PD21, the modeled mice were randomized into model, Wenyang, Jieyu, Wenyang Jieyu, and fluoxetine groups, with 15 mice in each group. The sucrose preference, tail suspension, and open field tests were carried out to evaluate the anxiety and depression-like behavior in mice. Enzyme-linked immunosorbent assay was used to measure the adrenocorticotrophic hormone (ACTH) and corticosterone (CORT) levels in mouse plasma. High performance liquid chromatography-electrochemical detector was used to determine the content of monoamine neurotransmitters in the hippocampus. Real-time fluorescence quantitative polymerase chain reaction was employed to determine the mRNA levels of genes in the 5-hydroxytryptamine (5-HT) system, hypothalamic-pituitary-adrenal (HPA) axis, and brain-derived neurotrophic factor (BDNF) signaling pathway in the hippocampus. Immunohistochemistry was employed to determine the expression levels of proteins in the 5-HT system and HPA axis in the hippocampus. The Simple Western system was used to determine the protein levels of BDNF and tyrosine kinase receptor B (TrkB) in the hippocampus. ResultCompared with the NC group, the model group exhibited depression-like behavior, which was significantly relieved by Wenyang Jieyu prescription and fluoxetine. Compared with the NC group, the model group showed elevated levels of CORT and ACTH in the plasma (P<0.01), which, however, were lowered by Wenyang Jieyu prescription and fluoxetine (P<0.05, P<0.01). Compared with the NC group, the model group showed inhibited expression of neurotransmitters in the hippocampus (P<0.05, P<0.01), while Wenyang Jieyu prescription and fluoxetine restored the expression of neurotransmitters (P<0.05, P<0.01). Compared with NC group, the model group showed inhibition of the 5-HTergic nerve and abnormal activation of the HPA axis, and Wenyang Jieyu prescription and fluoxetine regulated the abnormal state of the 5-HTergic nerve and HPA axis. Compared with NC group, the modeling down-regulated the mRNA and protein levels of BDNF and TrkB in the hippocampus (P<0.05, P<0.01), which, however, were recovered in Wenyang, Jieyu, Wenyang Jieyu, and fluoxetine groups (P<0.05, P<0.01). ConclusionThe mouse model of depression induced by MS+RS may present the syndrome of Yang deficiency and liver depression. Wenyang Jieyu prescription may increase the content of hippocampal neurotransmitters by regulating the 5-HT system and the BDNF signaling pathway mediated by the HPA axis, thereby alleviating depression-like behavior in mice.