Salidroside Regulates the miRNA-1343-3p/MAP3K6/MMP24 Signaling Molecules to Inhibit Proliferation and Invasion of Gastric Cancer Cells / 中山大学学报(医学科学版)

ObjectiveThe aim of this study is to investigate the role of salidroside in regulating the miR-1343-3p/MAP3K6 （mitogen-activated protein kinase kinase kinase 6）/MMP24 （membrane-type matrix metalloproteinase 24） signaling pathway to inhibit gastric cancer cell proliferation and migration. MethodsHuman gastric cancer cells （MGC-803） were divided into several groups based on different salidroside concentrations： a control group （0 μmol/mL）， a low-dose group （6 μmol/mL）， a medium-dose group （12 μmol/mL）， and a high-dose group （24 μmol/mL）. The anti proliferative effects of salidroside on human gastric cancer cells were evaluated by CCK-8 assay. Clonogenic assay was used to examine the effects of salidroside drugs on the clonogenic ability of human gastric cancer cells. Transwell assay was performed to detect the effect of salidroside on the invasive ability of human gastric cancer cells. Cell scratch assay was performed to detect the effect of salidroside on the migration ability of human gastric cancer cells. The miRNA expression profile was analyzed by using RNA-seq in cancer cells for 24 h after salidroside treatment. The differentially expressed miRNAs were clustered and their target genes were predicted. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） were used to analyze and predict the functions of these target genes， and the interaction networks were established. Immunocytofluorescence was used to detect the expression of target proteins， and the transcription of candidate genes was detected by q-PCR. ResultsCCK-8 cytotoxicity experiments showed that salidroside inhibited the proliferation of MGC-803 cells （P < 0.01）. Cell cloning experiments showed that salidroside reduced the clonal formation capacity of MGC-803 cells （P < 0.000 1）. Cell invasion experiments showed that salidroside reduced the MGC-803 cell invasion capacity （P < 0.000 1）. Cell scratch experiments showed that salidroside reduced the cell migration capacity （P < 0.000 1）. RNA-seq findings showed that the expression of 44 miRNAs changed significantly after salidroside treatment in cancer cells （P < 0.05）. Bioinformatic analysis showed that there were 1 384 target mRNAs corresponding to the differentially expressed miRNAs， and the expression of the tumor suppressor miR-1343-3p was significantly upregulated after salidroside treatment （P < 0.01），and resulted in down-regulated transcription of MAP3K6 and MMP24 genes which are related to the proliferation and migration of cancer cells （P < 0.05）. Immunofluorescence experiments demonstrated that salidroside reduced protein expression levels in MAP3K6 and MMP24 genes （P < 0.000 1）. q-PCR experiments showed that salidroside reduced the mRNA expression level of MAP3K6 and MMP24 genes （P < 0.000 1）， while miRNA expression in miR-1343-3p gene was upregulated （P < 0.000 1）. ConclusionSalidroside regulates the miRNA-1343-3p/MAP3K6/MMP24 signaling molecules to inhibit proliferation and invasion of gastric cancer cells.

miR-335 targets ENPP4 and FADS1 genes to mediate obesity in high-fat mice / 中华内分泌代谢杂志

Objective:To detect the differential expression of microRNA(miRNA) in abdominal fat of mice fed with high fat and to explore the role of highly expressed miR-335 in lipid metabolism.Methods:MicroRNA microarray was used to detect the differential expression of miRNAs in abdominal fat of mice fed with high fat. The target genes of miR-335 were predicted by Targetscan, the target genes of ectonucleotide pyrophosphatase/phosphordiester-ase 4(ENPP4) and fatty acid desaturase1(FADS1) were verified with double luciferase reporter system and point mutation test. miR-335 mimics was transfected into 3T3L1 cells to detect the expression of target gene mRNA and protein; Realtime PCR was used to detect the expression levels of miR-335 and target genes in different tissues of mice.Results:After high-fat feeding, the size of mice in the model group was significantly larger than the control group, and the serum total cholesterol and triglyceride levels of mice were significantly increased( P=0.016, P=0.014). miRNAs were differentially expressed in adipose tissues of mice fed with high fat, and the expression of miR-335 increased significantly( P=0.024). Double luciferase reporter system showed that miR-335 combined with the predicted target sites of ENPP4 and FADS1 through the " seed region" to inhibit the expression of ENPP4 and FADS1 genes, and the point mutation test confirmed the binding target sites between miR-335 and ENPP4/FADS1. MiR-335 mimics were transfected into 3T3L1 cells, the expression level of FADS1 mRNA decreased significantly( P=0.038), and the protein levels of ENPP4 and FADS1 decreased significantly( P=0.033, P=0.007). Realtime PCR showed that, miR-335 was significantly higher expressed in liver, brown adipose tissue, and brain after high-fat feeding, especially in white adipose tissue( P=0.002). The expression of ENPP4 and FADS1 in brain( P=0.006, P=0.034) and brown adipose tissue( P=0.014, P=0.037) decreased significantly, and the expression of ENPP4 in liver also decreased significantly after high-fat diet( P<0.001). Conclusion:miR-335 is a miRNA related to lipid metabolism and fat deposition. ENPP4 and FADS1 are the target genes of miR-335.

Up-regulated miR-452-5p suppressed the expression of RORα and promoted the proliferation and migration of HCC cells / 中国药科大学学报

@#To study the prognosis-related regulation mechanism of miR-452-5p and its influence on the proliferation and migration of hepatocellular carcinoma (HCC) cells, liver hepatocellular carcinoma (LIHC) dataset in The Cancer Genome Atlas (TCGA) was used to validate the differential expression of miR-452-5p and perform the Kaplan-Meier analysis of overall survival (OS).Target genes of miR-452-5p from TargetscanHuman and miRDB databases were predictived; and differentially expressed genes(DEGs) and weighted gene co-expression network analysis (WCGNA) were completed with GSE14520.Lipofectmine-2000 was used to transfect miR-452-5p mimics, mimics negative control, miR-452-5p inhibitor and inhibitor negative control into Huh7 cells,respectively.The mRNA and protein expression level of RORα in 4 groups were determined by RT-qPCR and Western blot.CCK-8 assay and Transwell assay were conducted to testify the capabilities of proliferation and migration.The regulation between miR-452-5p and RORα was confirmed by the dual luciferase reporter assay.After analysis in the TCGA-LIHC dataset, miR-452-5p had higher expression in HCC tissue than that in normal tissue, which was also associated with a shorter OS.RORα and LAMC1 were discriminated by intersecting of DEGs, WGCNA module genes, and predictive target genes.Survival analysis exhibited that dysregulation of RORα was significantly related to the OS.Overexpression of miR-452-5p in HCC cells suppressed the expression of RORα both in mRNA and protein, and also enhanced the viability and migration of HCC cells.The results of the dual luciferase reporter assay showed that miR-452-5p targeted 3′UTR of RORα.Up-regulated miR-452-5p inhibited the expression of RORα, facilitated the proliferation and migration of HCC cells, and promoted the progression and poor prognosis of HCC.

Differential expression profiles of miRNAs and their putative targets in Schistosoma mansoni during its life cycle

BACKGROUND Schistosomiasis is a disease caused by Schistosoma. Due to its complex life cycle, evolutionary position and sexual dimorphism, schistosomes have several mechanisms of gene regulation. MicroRNAs (miRNAs) are short endogenous RNAs that regulate gene expression at the post-transcriptional level by targeting mRNA transcripts. OBJECTIVES Here, we tested 12 miRNAs and identified their putative targets using a computational approach. METHODS We performed the expression profiles of a set of miRNAs and their putative targets during the parasite's life cycle by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). FINDINGS Our results showed differential expression patterns of the mature miRNAs sma-miR-250; sma-miR-92a; sma-miR-new_4-3p; sma-miR-new_4-5p; sma-miR-new_5-5p; sma-miR-new_12-5p; sma-miR-new_13-3p and sma-miR-new_13-5p. Interestingly, many of the putative target genes are linked to oxidative phosphorylation and are up-regulated in adult-worms, which led us to suggest that miRNAs might play important roles in the post-transcriptional regulation of genes related to energetic metabolism inversion during parasite development. It is noteworthy that the expression of sma-miR-new_13-3p exhibited a negative correlation on SmNADH:ubiquinone oxidoreductase complex I. MAIN CONCLUSIONS Our analysis revealed putative miRNA genes related to important biological processes, such as transforming growth factor beta (TGF-β) signaling, proteasome regulation, glucose and lipid metabolism, immune system evasion and transcriptional regulation.

Screening and preliminary analysis of differentially expressed miRNAs in gastric cancer cell-derived exosomes induced by Helicobacter pylori / 中华微生物学和免疫学杂志

Objective:To investigate the differentially expressed microRNAs (miRNAs) in human gastric carcinoma SGC-7901 cell-derived exosomes induced by Helicobacter pylori ( H. pylori), providing new clues for further elucidating the carcinogenic mechanism of H. pylori. Methods:Ultracentrifugation and exosome extraction kit were used to extract the exosomes released by the H. pylori-stimulated and negative control group, and transmission electron microscope(TEM), nanoparticle tracking analysis(NTA) and Western blot experiments were employed to identify exosomes. Then, exosomes were labeled with the fluorescent dye PKH67 and co-cultured with THP-1-derived macrophages. The internalization of exosomes by macrophages was observed by laser confocal fluorescent microscopy. Additionally, miRNA microarray chips were performed to detect the differentially expressed miRNAs of exosomes from the two groups of cells. Real-time fluorescence quantitative PCR (qRT-PCR) was used to verify the expression of four differentially expressed miRNAs. Furthermore, the target genes and their functions as well as the possible signal pathways involved of partial differentially expressed miRNAs were predicted and analyzed by bioinformatics software. Differentially expressed miR-382-5p was labeled by Cy3 to observe whether it could be transferred to macrophages through exosomes. The expression of phenotype molecule CD206 and the cytokines TNF-α, IL-6 and IL-10 in miR-382-5p mimic-transfected macrophages were analyzed by qRT-PCR and ELISA, and the proportion of cells expressing CD206 and HLA-DR was analyzed by flow cytometry. Results:The extracted exosomes were consistent with exosome morphology and highly expressed the surface marker proteins CD9, CD63 and TSG101. After co-culturing with THP-1 derived macrophages for 12 h, the exosomes could be internalized by macrophages. Compared with the control group, there were 130 up-regulated miRNAs and 111 down-regulated miRNAs in the H. pylori-stimulated group. Bioinformatic analysis showed that the potential target genes of partial differentially expressed miRNAs were mainly involved in the regulation of PI3K-AKT, NF-κB, JAK-STAT, stem cell pluripotency and other inflammation and tumor-related pathways. miR-382-5p could be transferred to macrophages through exosomes, and induced the expression of M2-type phenotype molecule CD206 and cytokines IL-10 in macrophages, while inhibited the expression of TNF-α and IL-6 and increased the proportion of CD206 high HLA-DR low cells. Conclusions:H. pylori treatment caused a significant change in the expression level of exosome miRNAs in SGC-7901 cells. Bioinformatics analysis demonstrated that the prospective targets of these differentially expressed miRNAs might play an important role in the regulation of inflammation and tumor-related signaling pathways. miR-382-5p might induce the M2-type polarization of macrophages.

Bioinformatics analysis of differential expression genes and microRNA-messenger RNA regulatory network in biliary atresia / 中华实用儿科临床杂志

Objective:To mining differential expression genes (DEGs) and establish a regulatory network of dysregulated microRNAs (miRNAs) and messenger RNAs (mRNAs) in biliary atresia (BA) spectrum via bioinforma-tics analysis, and to explore the pathogenesis of BA.Methods:GSE46960 dataset was download from gene expression omnibus (GEO). DEGs between normal liver tissues and BA tissues were analyzed using the GEO2R analysis tool.The functional and pathway enrichment analyses of DEGs were performed utilizing the Database for Annotation, Visualization and Integrated Discovery (DAVID6.8). A protein-protein interaction (PPI) network was constructed using the PPI database (STRING11.0) and Cytoscape_v3.7.1 software, and thus key genes were analyzed.BA-related miRNAs were obtained using the human miRNA disease database (HMDD_V3.0) and target mRNAs were predicted by the miRNA target prediction database (miRDB). The intersection between the predicted target mRNAs and DEGs from the GSE46960 dataset was selected.The regulatory network of miRNA-mRNA was constructed using Cytoscape software.Results:A total of 565 DEGs, including 352 up-regulated ones and 213 down-regulated ones were identified.Among them, up-regulated DEGs were enriched in extracellular matrix(ECM)-receptor interaction, focal adhesion kinase (FAK), Amoebiasis, and the phosphoinositide 3 kinase/protein kinase B(PI3K/Akt) pathway.Down-regulated DEGs were enriched in metabolic signaling, biosynthesis of antibiotics and steroid biosynthesis pathway.From the PPI network, 10 key genes were screened out.A complex miRNA-mRNA regulatory network was constructed based on screened DEGs.Conclusions:Identified DEGs and miRNA-mRNA regulatory network constructed in this study may help clarify the molecular mechanisms of BA.This study provides a new direction to explore promising molecular targets for the diagnosis and treatment of BA.

Differential expression profile of circRNAs in oral leukoplakia / 口腔疾病防治

Objective@#To investigate the differences and clinical significance of circRNA expression profiles in oral leukoplakia (OLK) tissues and normal oral mucosal (NOM) tissues.@*Methods@# High-throughput sequencing was used to detect differentially expressed circRNAs in 6 pairs of OLK and NOM tissues, and qRT-PCR was used to verify the expression of 10 circRNAs screened in 6 pairs of OLK and NOM tissues. The ring formation of circRNA was verified by RNase R digestion and Sanger sequencing, and the target circHLA-C was further verified by qRT-PCR in 20 pairs of OLK and NOM tissues. CircHLA-C was visualized using the UCSC genome browser (genome.ucsc.edu). The function of differentially expressed circRNAs was analyzed by GO and KEGG enrichment analyses. TargetScan and miRanda predicted the downstream miRNAs and mRNAs of the target circRNAs, and a ceRNA network related to the identified circRNAs was constructed in Cytoscape.@* Results@#Sequencing analysis showed that 366 circRNAs were significantly differentially expressed in OLK tissues, including 65 upregulated and 301 downregulated circRNAs. After qRT-PCR verification, 7 of the 10 screened circRNAs were expressed consistent with the sequencing results. The upregulated circHLA-C was confirmed to be a real circRNA with back-splice junction sites by RNase R digestion and Sanger sequencing. Correlation analysis showed a positive correlation between circHLA-C and the degree of OLK dysplasia. ROC curve analysis suggested that circHLA-C had potential value in diagnosing OLK with high accuracy and specificity.@*Conclusion@#CircRNA was significantly abnormally expressed in OLK tissues, and the upregulation of circHLA-C may be related to the degree of OLK dysplasia, providing guiding value for the diagnosis of OLK in the future.

Differential expression of circular RNA in patients with white matter hyperintensity and cognitive impairment / 中南大学学报(医学版)

OBJECTIVES@#White matter hyperintensity (WMH) is an important factor leading to cognitive impairment, and the mechanism has not been clarified. In recent years, studies have found that circular RNA (circRNA) has differential expression in cerebrovascular diseases. This study aims to analyze the expression profile of circRNA in peripheral blood mononuclear cell (PBMC) of patients with WMH with cognitive impairment, to screen the differentially expressed circRNA, and to explore the possible role of circRNA in WMH with cognitive impairment.@*METHODS@#CircRNA microarray was used to detect the circRNA expression profile of PBMC in patients with WMH with cognitive impairment, and in patients with WMH without cognitive impairment as well as in normal controls (3 cases each, male to female ratio of 2꞉1). The differentially expressed circRNA in patients with WMH with cognitive impairment was screened. The screening criteria for differentially expressed circRNA was fold change (FC) ≥2.0 (|log@*RESULTS@#Compared with the control group, there were 5 significantly up-regulated circRNA and 3 down-regulated circRNA in the WMH with cognitive impairment group; 8 circRNA were significantly up-regulated and 2 were down-regulated in the WMH without cognitive impairment group. When compared with the WMH with cognitive impairment group, no co-differentially expressed circRNA was found in WMH without cognitive impairment group and control group. Compared with the control group, the expression of hsa_circ_0092222 was up-regulated and the expressions of hsa_circ_0000662 and hsa_circ_0083773 were down-regulated in the WMH with cognitive impairment group and the WMH without cognitive impairment group, and there was no significant difference between the 2 groups (all @*CONCLUSIONS@#The circRNA expression profile of patients with WMH is changed significantly. The differentially expressed circRNA may be the cause of WMH; Hsa_circ_0092222, hsa_circ_0000662, and hsa_circ_0083773 may regulate the expression of target genes by targeting adsorption of the target miRNA, leading to brain white matter damage through Janus kinase 2 (JAK2)/signal transducers and activators of transcription (STAT3) signal pathway and Wnt signal pathway.There is no significant difference in circRNA expression profile between WMH with or without cognitive impairment. Cognitive impairment in patients with WMH may have other reasons.

Integration-based co-expression network analysis to investigate tumor-associated modules across three cancer types / 生物工程学报

In case/control gene expression data, differential expression (DE) represents changes in gene expression levels across various biological conditions, whereas differential co-expression (DC) represents an alteration of correlation coefficients between gene pairs. Both DC and DE genes have been studied extensively in human diseases. However, effective approaches for integrating DC-DE analyses are lacking. Here, we report a novel analytical framework named DC&DEmodule for integrating DC and DE analyses and combining information from multiple case/control expression datasets to identify disease-related gene co-expression modules. This includes activated modules (gaining co-expression and up-regulated in disease) and dysfunctional modules (losing co-expression and down-regulated in disease). By applying this framework to microarray data associated with liver, gastric and colon cancer, we identified two, five and two activated modules and five, five and one dysfunctional module(s), respectively. Compared with the other methods, pathway enrichment analysis demonstrated the superior sensitivity of our method in detecting both known cancer-related pathways and those not previously reported. Moreover, we identified 17, 69, and 11 module hub genes that were activated in three cancers, which included 53 known and three novel cancer prognostic markers. Random forest classifiers trained by the hub genes showed an average of 93% accuracy in differentiating tumor and adjacent normal samples in the TCGA and GEO database. Comparison of the three cancers provided new insights into common and tissue-specific cancer mechanisms. A series of evaluations demonstrated the framework is capable of integrating the rapidly accumulated expression data and facilitating the discovery of dysregulated processes.

Preliminary screening and analysis of differentially expressed circRNAs in esophageal squamous cell carcinoma / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To construct a circRNA profile of esophageal squamous cell carcinoma (ESCC) and analyze differentially expressed circRNAs. Methods: Samples were taken from 3 patients with esophageal squamous cell carcinoma who were hospitalized in the Department of Thoracic Surgery, Jintan Hospital, Jiangsu University from June 2018 to February 2019. The circRNA expression profile was constructed by high-throughput sequencing technique, and the circRNA differentially expressed in 3 pairs of esophageal squamous cell carcinoma tissues and adjacent tissues was detected. The biological functions and related signal pathways of these circRNA were analyzed by GO and KEGG techniques. Results: By comparing the expression levels of circRNA between esophageal squamous cell carcinoma and adjacent tissues, 905 differentially expressed circRNA were found, of which 404 were up-regulated and 501 were down-regulated. hsa_circ_0004390 was the CIRC RNA with the highest up-regulation factor (FC=7.9712), and novel_circ_0012687 was the one with the highest down-regulation factor. GO and KEGG analysis showed that these circRNA may be involved in biological processes such as cell cycle, cell components and protein binding of cancer cells, and signal pathways such as Hippo and cGMP-PKG. Conclusion: The expression profile analysis of circRNA in esophageal squamous cell carcinoma showed that the significantly differentially expressed circRNA could be used as a potential biomarker of esophageal squamous cell carcinoma.

Molecular Signatures of MicroRNA in Obesity Associated Obstructive Sleep Apnea in Middle-Aged Adults

Objective: Obstructive sleep apnea (OSA) is a respiratory disorder with multiple clinical outcomes and is now being recognized as a serious health concern across the globe. However, the mechanisms of its pathophysiology are still elusive. Also, to date, evidence of miRNA regulation of sleep apnea in the Indian sub-population is unknown. Methods: In this study, we investigated the expression pattern of certain potential obesity-linked miRNAs in OSA subjects. Seventy adult subjects (20 obese OSA, 20 non-obese OSA and 30 healthy) were selected for this study. Total RNA was extracted and the expression of miR-21, miR-27, miR-29 and let-7 (normalized with internal control RNU48) was analyzed by SYBR Green-based qPCR. Results: We selected miR-21, miR-27, miR-29 and let-7 for their documented role in obesity. Relative miRNA expression profiles revealed differential expressions of all four above mentioned miRNAs in both obese and non-obese OSA subjects compared to healthy controls. Statistical analysis revealed a significant correlation between miRNA expression with obesity-associated parameters in OSA subjects. Conclusion: Our study demonstrates the involvement of four miRNAs (miR-21, miR-27, miR-29 and let-7) as potential molecular players of obesity-associated OSA. Identification of miRNA targets and in-depth analysis of their molecular mechanism in disease pathogenesis is further warranted.

Expression changes of miRNA-29b-3p and miRNA-34c-3p in lung tissue of rats exposed to silica and A549 cells / 中华劳动卫生职业病杂志

Objective@#To investigate the role of microRNA-29b-3p (miRNA-29b-3p) and miRNA-34c-3p in the process of pulmonary fibrosis, we detected the expression levels of miRNA-29b-3p and miRNA-34c-3p in the lung tissue of rats exposed to silica and A549 cells.@*Methods@#SPF male Wistar rats were randomly divided into 1, 7, 14, 21, 28 d control group and silica (SiO2) dusting group, with 6 rats in each group. One-time non-exposure method was used to infuse 1ml SiO2 suspension. The rat SiO2 dusting group was established in the liquid, and the control rats were intratracheally injected with 1 ml of sterile physiological saline in the same manner. The lung tissues of each group were collected at the corresponding time points after dusting. Three of the rats were taken out for pathological observation, and the other three were used to screen differentially expressed miRNAs in lung tissue by miRNA microarray technology. A549 cells were cultured at the in vitro cell level and divided into control group, SiO2 stimulation group and TGF-β1 stimulation group, and cells were collected at 12, 24 and 48 h after treatment. The expression levels of miRNA-29b-3p and miRNA-34c-3p in rat lung tissue and A549 cells were verified by real-time PCR (qRT-PCR), target gene prediction of miRNA-29b-3p and miRNA-34c-3p and perform GO enrichment analysis and KEGG pathway analysis.@*Results@#The weight growth rate of the control group was significantly higher than that of the SiO2 dusting group. Compared with the control group, the lung mass and lung coefficient of the SiO2 dusting group were significantly increased (P<0.05). The inflammatory response of the lungs in the control group was significantly reduced at 21 and 28 days, and the inflammatory cells infiltrated in the lung tissue of the SiO2 group. The rats in the control group had a small amount of collagen at 21 and 28 days. A large amount of collagen fiber deposition began to appear in the lung tissue of rats exposed to SiO2 for 21 days. Compared with the control group, the expression levels of miRNA-29b-3p and miRNA-34c-3p in the SiO2 dusting group were significantly down-regulated, and there was significant difference compared with the control group (P<0.05). The expression levels of miRNA-29b-3p and miRNA-34c-3p in A549 cells treated with SiO2 and human recombinant TGF-β1 were significantly lower than those in the control group at 24 h and 48 h, and the difference was statistically significant (P<0.05).@*Conclusion@#Down-regulation of miRNA-29b-3p and miR-34c-3p in rat lung tissue A549 cells may be associated with the development of early silicosis and is expected to be an indicator of early silicosis diagnosis and prognosis.

Preliminary analysis of differential expression of miRNA-423-5p and miRNA-26a-5p in lung tissue of early silicotic rats / 中华劳动卫生职业病杂志

Objective@#To screen the changes of microRNA (miRNA) expression profiles in lung tissues of early silicosis rats, and provide a basis for functional analysis of differential microRNA.@*Methods@#SPF Wistar male rats were randomly divided into a negative control group and SiO2-exposed groups, with 30 rats in each group. The model of silicosis in rats was established by intratracheal instillation of 1 ml SiO2 suspension, and the control rats were treated with 1mL in the same way to sterilize normal saline. The lung tissues of two group were collected at the 1, 7, 14, 21, 28 d after SiO2-exposed. Three of the rat lung tissues were used for pathological observation, and the other three were used to screen differentially expressed miRNAs in lung tissue by miRNA microarray technology. miRNA chip screening and RT-qPCR were used to verify the expression levels of miRNA-423-5p and miRNA-26a-5p in the two groups. miRNA-423-5p and miRNA-26a-5p are predicted by target genes and analyzed by GO (gene ontology) enrichment analysis and KEGG (kyoto encyclopedia of genes and genomes) pathway analysis.@*Results@#In the control group, the inflammatory response of lung tissue 21 and 28 days was significantly reduced compared with 1, 7 and 14 days, and the inflammatory cells infiltrated in the lung tissue of the SiO2-exposed rats. The rats in the control group had a small amount of collagen at 21 and 28 days, but a large amount of collagen fiber deposition began to appear in the lung tissue of rats exposed to SiO2 after 21 days. Compared with the control group, the expression levels of micro RNA-423-5p was significantly up-regulated and the expression of microRNA-26a-5p was significantly down-regulated in the SiO2-exposed rats lung tissues dust at different time points (P<0.05) .@*Conclusion@#The up-regulation of miRNA-423-5p and the down-regulation of miRNA-26a-5p in lung tissues of early silicotic rats may be related to the occurrence and development of early silicosis.

Bioinformatics Screening of Key Genes and Candidate Therapeutic Drugs of Osteoarthritis / 中国实验方剂学杂志

Objective: To explore the key genes and potential therapeutic drugs for osteoarthritis (OA) by bioinformatics.Method: The microarray data GSE55235 was downloaded from the data platform of gene expression omnibus (GEO) and the differentially expressed genes were screened by R language software (3.5.0).Then,the differentially expressed genes were subjected to gene ontology (GO) enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) signaling pathway analysis with David online database.The protein-protein interaction was analyzed by String 10.5 online database and visual editing was analyzed by Cytoscape v3.6.1 software.Subnetwork module analysis was utilized by MCODE plugin to screen the core genes in the process of OA.Finally,small molecule drugs with potential treatment for OA were analyzed by connectivity map (CMap) database.Result: A total of 556 differentially expressed genes were screened,among which 252 were up-regulated and 304 were down-regulated.These genes were mainly involved in extracellular matrix (ECM) organization,inflammatory response,cell adhesion,immune response,collagen binding,etc.The analysis of KEGG pathway showed that differential genes were mainly involved in ECM-receptor interaction,phosphatidylinositol 3 kinase-protein kinase B (PI3K/Akt) signaling pathway and osteoclast differentiation.Some genes,such as interleukin-6(IL-6),JUN,vascular endothelial growth factor α(VEGFA),FOS,MYC and early growth response gene-1(EGR-1),activating transcription factor-3(ATF-3),playing critical role in the process of OA were identified by protein-protein interaction.Some potential small molecular drugs for the treatment of OA have also been screened,such as lycorine and anisomycin.Conclusion: The selected key genes may be targets for the diagnosis of OA or potential targets for the treatment of OA,and the selected small molecular drugs can be developed as the key drugs for the treatment of OA.

The alteration of lncRNA expression in peripheral blood mononuclear cells and the effects of anti-anxiety treatment on lncRNA expression of generalized anxiety disorder patients / 中华行为医学与脑科学杂志

Objective@#To confirm expression alteration of long non-coding RNA(lncRNA) in peripheral blood mononuclear cells (PBMCs) of generalized anxiety disorder(GAD) patients and anti-anxiety treatment effects on aberrant expression of lncRNAs.@*Methods@#Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was employed in 80 GAD patients and 40 healthy participants to confirm 10 aberrant lncRNAs screened by microarray expression profiling.And 26 out of all the 80 GAD patients were recruited for lncRNA expression level testing and Hamilton Anxiety Scale(HAMA) assessments before and after 6 weeks’ treatment.@*Results@#Six of ten lncRNAs selected by array profiling (lncRNA4(7.44±2.26), lncRNA5(6.83±2.28), lncRNA6(8.09±2.30), lncRNA8(9.10±2.36), lncRNA9(7.66±2.12), lncRNA10(7.34±2.12)) were verified by qRT-PCR that the lncRNA expression levels were significantly up regulated in GAD patients compared with healthy controls(Z=-3.022--1.996, P<0.05 or 0.01), and lncRNA4(9.73±2.53), lncRNA6(9.91±2.01), lncRNA8(10.48±1.68), lncRNA9(9.02±1.58), lncRNA10(9.04±2.08) were down regulated significantly after 6 weeks’ anti-anxiety treatment(Z=-3.180--2.530, P<0.05 or 0.01) along with signicant reduction of total HAMA score (11.19±8.37), dimension scores of somatic anxiety(5.31±4.76), psychic anxiety(5.88±3.82) (t=5.502-5.971, P<0.01). The alterations of lncRNA4, lncRNA6, lncRNA8, lncRNA9, lncRNA10 were positively correlated with that of HAMA total score and psychic anxiety score(r=0.39-0.69, P<0.05 or 0.01), and alteration of lncRNA6, lncRNA8, lncRNA10 had positive correlation with that of somatic anxiety score(r=0.44-0.59, P<0.01).@*Conclusion@#The expression level of lncRNA4, lncRNA5, lncRNA6, lncRNA8, lncRNA9, lncRNA10 are up-regulation in PBMCs of GAD patients and anti-anxiety treatment can reverse the expression level of lncRNAs. Alteration of lncRNA expression has osculatory association with improvement of anxious symptom.

Differential expression of 14-3-3 gene in different stages of Taenia solium / 中华地方病学杂志

Objective To investigate the expression difference of 14-3-3 gene in different stages of Taenia solium (Ts),and to provide basic information for exploring the regulation mechanism of Ts14-3-3 gene in growth and development of Ts.Methods Adult worms were collected from patients with taeniasis solium in the taeniasis epidemic area of Yajiang County,Ganzi Tibetan Autonomous Prefecture,Sichuan Province.After determining the species through morphological observation under the light microscope and transcribed spacer 1 (ITS1) method,it was identified as Ts,then piglets were infected to obtain the Cysticercus cellulosae of Ts (larvae).Reverse transcription PCR was used to detect the expression of Ts14-3-3 gene in the adult stage and larval stage of Ts,and the relative expression levels of each were detected by real-time fluorescent quantitative PCR.Results Under light microscope,it could be seen that the collected adult worm scolex consisted of 4 suckers,rostellum and hooks,which was consistent with the typical characteristics of the Ts scolex;the worm sample could amplify the target fragment which was consistent with the length of the ITS1 gene by reverse transcription PCR,and was determined to be Ts.The Ts14-3-3 gene family members Ts14-3-3.1,Ts14-3-3.2,Ts14-3-3.3,Ts14-3-3.4,Ts14-3-3.5 and Ts14-3-3.6 were expressed in the Ts adult stage and larval stage.Compared with the larval stage,the expression levels of Ts14-3-3.1 (0.47 ± 0.09 vs 1.01 ± 0.23),Ts14-3-3.2 (0.31 ± 0.09 vs 1.05 ± 0.14),Ts14-3-3.3 (0.64 ± 0.23 vs 1.26 ± 0.23) and Ts14-3-3.4 (0.30 ± 0.09 vs 0.79 ± 0.23) were significantly decreased in the adult stage (t =-3.816,-7.093,-3.377,-3.481,P ＜ 0.01 or ＜ 0.05);the expression levels of Ts14-3-3.5 (3.59 ± 0.09 vs 0.99 ± 0.12) and Ts14-3-3.6 (5.74 ± 2.76 vs 1.03 ± 0.60) were significantly increased (t =30.714,9.718,P ＜ 0.01).Conclusion The expression of Ts14-3-3 gene has stage specificity and the Ts14-3-3 gene may play a special regulatory role in different stages of Ts.

Study on the Effects of Danhong Injection on Gene Expression Profile of Acute Myocardial Infarction Model Rats by Gene Chip Technique / 中国药房

OBJECTIVE： To investigate the effects of Danhong injection （DHI） on gene expression profile of acute myocardial infarction （AMI） model rats. METHODS： Male SD rats were randomly divided into sham operation group， model group and DHI group （0.76 mL/kg）， with 10 rats in each group. AMI model was established by ligation of left anterior descending coronary artery in model group and DHI group. After modeling， sham operation group and model group were given constant volume of normal saline intramuscularly， and DHI group was given relevant medicine intramuscularly， once a day， for consecutive 14 days. After last administration， myocardial tissue in the marginal zone of infarction was separated. The change of gene expression profile was detected by gene chip technique. Using fold-change of relative expression as index， differentially expressed microRNA （miRNA） were screened. On the basis of retrieving their corresponding genes， gene ontology （GO） and KEGG pathway enrichment analysis were carried out by using DAVID bioinformatics resource database and KEGG pathway database， respectively. TargetScan database was used to predict the target gene messenger RNA （mRNA） corresponding to differentially expressed miRNA. Cytoscape 3.6.1 software was used to construct and analyze the miRNA-mRNA network. Agilent GeneSpring GX v11.5 software was used to screen target genes and miRNA related to inflammation in the above networks. RESULTS： Compared with sham operation group， there were 22 differentially expressed miRNAs in model group， 5 up-regulated and 17 down-regulated. Compared with model group， there were 26 differentially expressed miRNAs in DHI group， and all of them were up-regulated. The differentially expressed miRNAs related to DHI therapy for AMI included rno-let-7a-5p， rno-let-7d-5p， rno-let-7f-5p， rno-miR-26b-5p， rno-miR-29b-3p， cel-miR-39-3p， cel-miR-39-5p， rno-miR-142-5p， rno-miR-191a-5p， rno-miR-409a-3p. Results of GO analysis and KEGG pathway enrichment analysis showed that differentially expressed miRNAs were mainly concentrated in membrane-bound organelles， cytoplasm， endometrial system and other cell components. The molecular functions such as protein binding and ion binding were exerted through biological processes such as anatomical structure development， multicellular tissue development and development process，which were mainly enriched in calcium signaling pathway， PPAR signaling pathway， VEGF signaling pathway， cell apoptosis， glycosylphosphatidylinositol anchored biosynthesis， valine， leucine and isoleucine degradation， etc. miRNA-mRNA network analysis showed that there were 25 target gene mRNAs corresponding to differentially expressed miRNA and 24 miRNAs related to it. There were 6 inflammation-related target genes （IL6， IL1b， TNF， TLR4， CRP， CXCL12） in this network， involving 19 differentially expressed miRNAs. CONCLUSIONS： The therapeutic effect of DHI on AMI may be related to regulating the expression of related miRNA， affecting signal transduction of calcium ion， PPAR and VEGF pathways， and regulating the secretion of inflammatory markers such as interleukin， chemokine and C-reactive protein.

Synthesis of triterpenoid saponins from Aesculus chinensis based on transcriptome data / 中国中药杂志

Aesculus chinensis belongs to Hippocastanaceae family,bears medicinal and ornamental values. The oleanane type triterpenoid saponin aescin is regarded as active ingredient and accumulated in seed. In order to understand its molecular basis of the triterpenoid biosynthesis,we used high-throughput sequencing under Illumina Hi Seq 2000 platform to obtain the transcriptome data of seed and flower from A. chinensis to further mine the genes involved in its metabolic pathway. Unigene's de novo splicing was performed using Trinity software; the transcriptome results were annotated with KEGG database to predict the specific pathways of the aescin triterpenoid metabolism. Terpenoid and triterpenoid pathways were found from transcriptome data,and forty seven and twenty seven corresponding genes were uncovered respectively. It was found that there are eight kinds of enzymes related to the terpenoid metabolism pathway precursors and three kinds of enzymes related to the triterpenoid metabolism pathway. In this study,five genes corresponding to triterpene cyclase were analyzed in A. chinensis for the first time,which may participate in the synthesis of triterpenoid. It' s revealed that there were thirty three differential genes associated with the ko00900 and ko00909 pathways by analysis on the difference in transcriptome expression between seeds and flowers; seventeen unigenes were up-regulated and sixteen unigenes were down-regulated in the seeds relative to flowers. In this study, qRT-PCR experiments were used to verify the expression of three key enzyme genes of SQE( Unigene25806),HMGS( Unigene36710),and β-AS( Unigene33291). The results of qRT-PCR were consistent with the transcriptome data. The candidate genes related to triterpenoid saponin aescin synthesis in A. chinensis found in this study can provide theoretical basis for the metabolism synthesis and regulation of aescin.

Acute 1,2-dichloroethane exposure induced differential gene expression of liver phaseⅠmetabolic enzymes in SD rats / 中国职业医学

OBJECTIVE: To investigate the effect of 1,2-dichloroethane(1,2-DCE) acute inhalation exposure on the differential gene expression of phase Ⅰ metabolic enzymes. METHODS: The specific pathogen free SD rats were randomly divided into control group(16 rats), low-and high-dose groups(24 rats in each group, half males and half females). Low-and high-dose group were given daily 600, 1 800 mg/m~(3 ) of 1,2-DCE, and the control group given the fresh air by dynamic inhalation for 8 hours per day for consecutive 7 days. After the end of exposure, the relative mRNA expression of cytochrome P450 2 E1(CYP2 E1), alcohol dehydrogenase(ADH1) and acetaldehyde dehydrogenase 3 alpha 1(ALDH3α1) in the liver tissue was detected by real-time fluorescence quantitative polymerase chain reaction. RESULTS: The relative expression of CYP2 E1 in male high-dose group was higher than that in male low-dose group and female high-dose group(P<0.05). The relative expression of ADH1 in male low-and high-dose groups was higher than that in male control group(P<0.05). The relative expression of ADH1 in male high-dose group was higher than that in male low-dose group and female high-dose group(P<0.05). The relative expression of ALDH3α1 in high-dose group was higher than that in control group and low-dose group(P<0.05). CONCLUSION: High dose 1,2-DCE could increase the gene expression of phase Ⅰ metabolic enzymes in rat liver. The 1,2-DCE has more obvious effect in male rats than in female rats.

The differential expression of microRNA in human gastric cancer MGC803 cells induced by diallyl disulfide / 中国药理学通报

Aim: To investigate the expression and i-dentification of differential miRNAs in human gastric cancer MGC803 cells induced by diallyl disulfide (DADS). Methods: Differential miRNAs expression in human gastric cancer MGC803 cells induced by DADS was detected and identified by miRNA chip and qPCR. Results: MiRNAs chip detection showed upregulation of miR-200b, miR-22, miR-7, miR-143, miR-138, miR-34a and miR-150, and down-regulation of miR-222, miR-21, miR-15b, miR-182 and miR-18a in differential miRNAs of MGC803 cells treated with 3 0 mg · IT-1 DADS at 24 h(P<0.05). And qPCR demonstrated that the expressions of miR-200b, miR-22, miR-7, miR-143, miR-138, miR-34a and miR-150 was up-regulated in MGC803 cells treated with 30 mg · L-1 DADS(P <0. 05). Moreover, qPCR showed that the expressions of miR-200b and miR-22 in various human gastric cancer cells including MGC803, BGC823, MKN28, SGC7901 and HGC27 cells were lower than normal human gastric cancer GES-1 cells (P <0. 05). The expression of miR-200b and miR-22 in gastric cancer tissues was significantly lower than that in normal gastric tissues (P < 0. 05). Conclusions: The expression of down-regulation of 7 miRNA and up-regulation of 5 miRNA in differential miRNAs in MGC803 cells induced by DADS. The expression of down-regulation of miR-200b and miR-22 in gastric cancer tissues and cells. DADS could up-regulate the expression of miR-200b and miR-22 in gastric cancer cells.