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.

Advances in epigenetic regulation of Chinese hamster ovary cells / 生物工程学报

Chinese hamster ovary (CHO) cells play an irreplaceable role in biopharmaceuticals because the cells can be adapted to grow in suspension cultures and are capable of producing high quality biologics exhibiting human-like post-translational modifications. However, gene expression regulation such as transgene silencing and epigenetic modifications may reduce the recombinant protein production due to the decrease of expression stability of CHO cells. This paper summarized the role of epigenetic modifications in CHO cells, including DNA methylation, histone modification and miRNA, as well as their effects on gene expression regulation.

Promising roles of non-exosomal and exosomal non-coding RNAs in the regulatory mechanism and as diagnostic biomarkers in myocardial infarction / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Non-exosomal non-coding RNAs (non-exo-ncRNAs) and exosomal ncRNAs (exo-ncRNAs) have been associated with the pathological development of myocardial infarction (MI). Accordingly, this analytical review provides an overview of current MI studies on the role of plasma non-exo/exo-ncRNAs. We summarize the features and crucial roles of ncRNAs and reveal their novel biological correlations via bioinformatics analysis. The following contributions are made: (1) we comprehensively describe the expression profile, competing endogenous RNA (ceRNA) network, and "pre-necrotic" biomarkers of non-exo/exo-ncRNAs for MI; (2) functional enrichment analysis indicates that the target genes of ncRNAs are enriched in the regulation of apoptotic signaling pathway and cellular response to chemical stress, etc.; (3) we propose an updated and comprehensive view on the mechanisms, pathophysiology, and biomarker roles of non-exo/exo-ncRNAs in MI, thereby providing a theoretical basis for the clinical management of MI.

Research Progress of Exosomal microRNA in Cardiovascular Disease and Its Forensic Application Prospects / 法医学杂志

Exosomal microRNAs (miRNAs) are miRNAs that are mediated by exosomes to achieve cell-to-cell communication, and they are widespread in organisms. In recent years, the key role of the multiple biological functions of exosomal miRNAs in the occurrence and development of cardiovascular diseases has been confirmed by a large number of studies, which has become a hot spot in clinical and basic research. Sudden cardiac death caused by cardiovascular disease is one of the important contents in forensic medical identification. This article introduces the research progress of cardiovascular disease prediction, treatment and prognosis on exosomal miRNA. The prospects of the application in forensic medical identification are discussed.

Research progress of microRNA in proliferative vitreoretinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

@#Proliferative vitreoretinopathy(PVR)is a eye disease characterized by the formation of epiretinal membranes(ERM)composed of extracellular matrix(ECM)and various types of cells in the vitreous and/or the surface of the retina through the wound repair and fibrotic process. ERM shrinks to form retinal folds and stretches the retina to cause retinal detachment(RD). Epithelial-mesenchymal transition(EMT)of retinal pigment epithelial(RPE)cells and accumulation of ECM are considered to be the main pathological mechanisms for the formation of ERM. RPE cells undergo a process named EMT induced by transforming growth factor-β(TGF-β), by which differentiated epithelial cells go through epithelial phenotypic loss, the weakness of cell-cell contact and mesenchymal phenotype expression. Fibroblast-like cells differentiated from mesenchymal cells produce ECM and other components, which forms ERM together with glial cells and fibroblasts, <i>etc</i>. Recent studies indicated a lot of cytokines/growth factors, transcriptional factors, and microRNA(miRNA)regulate the development of EMT in RPE cells, in which miRNA is a novel and powerful regulatory gene and plays a critical regulatory role in the EMT process of PVR. This review focuses on the current understandings of the mechanism and the interventional treatments of miRNA in PVR.

LncRNA-m18as1 competitively binds with miR-18a-5p to regulate follicle-stimulating hormone secretion through the Smad2/3 pathway in rat primary pituitary cells / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Long noncoding RNAs (lncRNAs) are expressed in different species and different tissues, and perform different functions, but little is known about their involvement in the synthesis or secretion of follicle-stimulating hormone (FSH). In general, we have revealed lncRNA‍‒‍microRNA (miRNA)‍‒‍‍messenger RNA (mRNA) interactions that may play important roles in rat primary pituitary cells. In this study, a new lncRNA was identified for the first time. First, we analyzed the gene expression of lncRNA-m18as1 in different tissues and different stages by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and observed the localization of lncRNA-m18as1 with fluorescence in situ hybridization, which indicated that this lncRNA was distributed mainly in the cytoplasm. Next, we used RT-qPCR and enzyme-linked immunosorbent assay (ELISA) to analyze the regulation of FSH synthesis and secretion after overexpression or knockdown of lncRNA-m18as1 and found that lncRNA-m18as1 was positively correlated with FSH synthesis and secretion. In addition, mothers against decapentaplegic homolog 2 (Smad2) was highly expressed in our sequencing results. We also screened miR-18a-5p from our sequencing results as a miRNA that may bind to lncRNA-m18as1 and Smad2. We used RNA immunoprecipitation-qPCR (RIP-qPCR) and/or dual luciferase assays to confirm that lncRNA-m18as1 interacted with miR-18a-5p and miR-18a-5p interacted with Smad2. Fluorescence in situ hybridization (FISH) showed that lncRNA-m18as1 and miR-18a-5p were localized mainly in the cytoplasm. Finally, we determined the relationship among lncRNA-m18as1, miR-18a-5p, and the Smad2/3 pathway. Overall, we found that lncRNA-m18as1 acts as a molecular sponge of miR-18a-5p to regulate the synthesis and secretion of FSH through the Smad2/3 pathway.

Screening of IGF-1, NOS, FGF23 and Sclerost-Targeted Mechanoresponsive MicroRNAs / 医用生物力学

Objective To screen the secretory factor-related, mechanoresponsive microRNAs (miRNA) of osteocytes. Methods Cyclic mechanical tensile strain (ε=2.5,f=0.5 Hz) was applied to osteocytes and osteoblasts cultured in vitro respectively, and the differentially expressed miRNAs only in the osteocytes were screened out by using miRNA chip. Through bioinformatics technology, in these differentially expressed miRNAs, the target genes of secretory factors including insulin-like growth factor-1(IGF-1), nitric oxide synthesase (NOS), fibroblast growth factor 23 (FGF23) and sclerostin (SOST) were further screened out. Then the selected miRNAs were compared with the biochip detected, differentially expressed miRNAs in femur bone of the mice which were trained on treadmill, and four of these miRNAs were randomly selected for quantitative PCR verification. Results For the 77 differentially expressed miRNAs only in the mechanically strained osteocytes in vitro, 22 miRNAs whose target genes were the 4 secreted factors (IGF-1, NOS, FGF23 and SOST), were screened out. Moreover, a total of 11 miRNAs in the 22 miRNAs were differentially expressed in femur bone of the treadmill trained mice with the same trend as those in osteocytes in vitro, and the randomly selected miR-361-3p, miR-3082-5p, miR-6348 and miR-706 were confirmed to be differentially expressed with the same trend in femur bone and osteocytes. Conclusions These mechanoresponsive miRNAs differentially expressed only in osteocytes, such as miR-361-3p, miR-3082-5p, miR-6348 and miR-706, probably influence osteoblastic differentiation or bone metabolism through regulating the secretory factors.

Analysis of Neuroprotective Effect of Danggui Shaoyaosan on APPswe/PS1ΔE9 Transgenic Mice Based on Circular RNA Sequencing / 中国实验方剂学杂志

Objective:To investigate the neuroprotective effects of Danggui Shaoyaosan （DSS） on APP_swe/PS1_ΔE9transgenic （APP/PS1） mice and its mechanism related to circular RNA （circRNA）. Method:Totally twenty 6-month-old APP/PS1 mice were divided into model group and DSS group， and 10 C57BL/6 wild-type mice were set as the normal control group. The normal group and model group received the same volume of normal saline， and DSS group received drug by gavage administration， all for 8 weeks. The differentially expressed circRNA of APP/PS1 mice before and after DSS intervention was analyzed by circRNA sequencing to construct circRNA-miRNA mRNA interaction network. The results of cricRNA sequencing were then verified by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The protein expression levels of phosphoinositide 3-kinase （PI3K）， p-PI3K， protein kinase B1 （Akt1）， p-Akt1， B lymphocytoma-2 （Bcl-2）， and Bcl-2-Associated X protein （Bax） in the hippocampus were detected by immunoblotting （Western blot）. The protein expression of Caspase-3 in the hippocampus was detected by immunohistochemistry and the level of apoptosis in the hippocampus was detected by the TUNEL method. Result:Compared with the model group， there were 90 differentially expressed circRNA after intervention with DSS， of which 46 were up-regulated and 44 down-regulated. Compared with the normal group， the expression levels of circRNA1398 and circRNA1399 in the model group decreased， and the expression levels of miR-103-3p， miR-153-3p， miR-143-3p， and miR-143-5p increased. Compared with the model group， the expression levels of circRNA1398 and circRNA1399 in the DSS group were up-regulated， while the expression levels of miR-103-3p， miR-153-3p， miR-143-3p， and miR-143-5p were down-regulated. Compared with the normal group， the expression of p-PI3K， Akt1， p-Akt1， and Bcl-2 in the model group decreased （<italic>P</italic><0.05，<italic> P</italic><0.01）， and the expression of Bax and Caspase in the model group increased （<italic>P</italic><0.01）. Compared with the model group， the expression of p-PI3K， Akt1， p-Akt1， and Bcl-2 in the hippocampus of the DSS group increased （<italic>P</italic><0.01）， and the protein expression of Bax and Caspase decreased （<italic>P</italic><0.01）. Compared with the normal group， the apoptosis level in the hippocampus of the model group increased， with an apoptosis rate of （43.76±2.92）%. Compared with the model group， the apoptosis rate of DSS group was （24.64±3.39）%. Conclusion:DSS can activate PI3K/Akt pathway and inhibit apoptosis in hippocampal neurons of APP / PS1 mice， and play a neuroprotective role. The specific mechanism may be related to the regulation of circRNA1398 and circRNA1399 expression and the corresponding miRNA expression.

Bioinformatics analysis of miRNAs in mild cognitive impairment due to Alzheimer's disease / 上海交通大学学报（医学版）

Objective: To analyze the expression profile of plasma microRNA (miRNA) in patients with mild cognitive impairment (MCI) due to Alzheimer's disease (AD) by bioinformatics method, and explore its pathogenesis at the level of genetic regulation. Methods: Five MCI patients due to AD and five control participants were recruited. The plasma miRNA expression profiles were analyzed by miRNA microarray sequencing. Target genes of significantly up-regulated miRNAs were detected by TargetScan 7.2 database. The miRNA-gene interaction network of significantly up-regulated miRNAs was established by Cytoscape software, and the key miRNAs of the network were analyzed. The target genes of key miRNAs were analyzed by Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genome (KEGG) pathway analysis using R packages. Results: There are 13 up-regulated miRNAs in the plasma of MCI patients due to AD, and 5 of them were key miRNAs in miRNA-gene interaction network. Target genes of these miRNAs were mainly involved in biological process such as synaptic plasticity reg-ulation, Wnt signaling pathway, synaptic vesicle transport and synaptic vesicle localization, as well as Ras signaling pathway, mitogen-activated protein kinase (MAPK) signaling pathway and glycolysis/gluconeogenesis pathway. Conclusion: Five up-regulated miRNAs in plasma of MCI due to AD may be the main regulators involved in the pathological mechanism of AD, which can be used as potential biomarkers for diagnosis of MCI due to AD.

Differential MiRNA/mRNA Expression Profiling and Functional Network Analysis for MC3T3-E1 Cells with Microgravity Stimulation Based on RNA-seq / 医用生物力学

Objective To investigate the effect of microgravity on MC3T3-E1 osteoblast differentiation. Methods The differential miRNA and mRNA expression profiling of MC3T3-E1 cells during exposure to microgravity were established by RNA transcriptome sequencing technology (RNA-seq). The RNA sequencing results were validated using quantitative real-time polymerase chain reaction (q-PCR). Bioinformatic analyses were applied for further study of these differentially expressed miRNAs and mRNAs. Results Compared with control (CON) group, A total of 1 912 coding transcripts and 160 miRNAs were detected along with osteogenic differentiation in simulated microgravity (SMG) group. Bioinformatic analysis revealed 10 core regulatory genes including 7 mRNAs and 3 miRNAs. Based on the analysis and verification, one miRNA, miR-9_6666-5p, was identified, which might play an important role in osteogenic differentiation process under microgravity. Conclusions The process of osteoblast differentiation was repressed under microgravity which might be related to the changed expression profile of miRNA/mRNA. The research findings can contribute to the better understanding of the molecular mechanisms of mRNA and miRNAs in osteogenic differentiation and bone formation under the microgravity condition.

Screening of miRNAs associated with lymph node metastasis in Her-2-positive breast cancer and their relationship with prognosis / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The aim of this study was to identify some biomarkers for predicting lymph node metastasis and prognosis of human epidermal growth factor receptor 2 (Her-2)-positive breast cancer (BC). We analyzed correlations between microRNAs (miRNAs) and the prognosis of patients with BC based on data collected from The Cancer Genome Atlas (TCGA) database. The expression levels of miR-455, miR-143, and miR-99a were measured in clinical samples of Her-2-positive BC patients with different degrees of lymph node metastasis. We investigated the impacts of overexpressed miR-455 on the proliferation and invasiveness of MDA-MB-453 cells and measured its effects on the expression of long non-coding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) by quantitative real-time polymerase chain reaction (qRT-PCR). The expression of miR-455 was significantly and positively correlated to the prognosis and overall survival (OS) of the BC (P=0.028), according to TCGA information. The expression level of miR-455 was positively correlated with OS and relapse-free survival (RFS) of patients with Her-2-positive BC, and was negatively correlated with the number of metastatic lymph nodes (P<0.05). Transwell assay suggested that MDA-MB-453 cells became much less invasive (P<0.01) after being transfected with miR-455 mimics. During the qRT-PCR, the expression level of MALAT1 declined significantly after transfection (P<0.01). Overexpressed miR-455 significantly inhibited the proliferation and migration of MDA-MB-453 cells and the expression of MALAT1. We conclude that miR-455 may be a useful potential biomarker for forecasting lymph node metastasis and the prognosis of Her-2-positive BC patients. miR-455 may play an important role in lymph node metastasis of BC by interacting with MALAT1.

Circulating microrna expression profiles in patients with stable and unstable angina

OBJECTIVES: High incidence and case fatality of unstable angina (UA) is, to a large extent, a consequence of the lack of highly sensitive and specific non-invasive markers. Circulating microRNAs (miRNAs) have been widely recommended as potential biomarkers for numerous diseases. In the present study, we characterized distinctive miRNA expression profiles in patients with stable angina (SA), UA, and normal coronary arteries (NCA), and identified promising candidates for UA diagnosis. METHODS: Serum was collected from patients with SA, UA, and NCA who visited the Department of Cardiovascular Diseases of the Meizhou People's Hospital. Small RNA sequencing was carried out on an Illumina HiSeq 2500 platform. miRNA expression in different groups of patients was profiled and then confirmed based on that in an independent set of patients. Functions of differentially expressed miRNAs were predicted using gene ontology classification and Kyoto Encyclopedia of Genes and Genomes pathway analysis. RESULTS: Our results indicated that circulating miRNA expression profiles differed between SA, UA, and NCA patients. A total of 36 and 161 miRNAs were dysregulated in SA and UA patients, respectively. miRNA expression was validated by reverse transcription quantitative polymerase chain reaction. CONCLUSION: The results suggest that circulating miRNAs are potential biomarkers of UA.

Identification and characteristic analysis of enhancer-miRNA regulatory pairs in hepatic carcinoma / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To explore the regulatory relationship between enhancer and miRNA and the characteristics of the enhancers that regulate miRNA in hepatic carcinoma and normal hepatic tissues, and to screen the differentially expressed miRNAs regulated by enhancers as well as their association with the treatment targets in liver cancer. Methods: Based on the TCGA and FANTOM5 databases, Co-expression and 3D genomic analysis were performed on 417 samples of enhancers and miRNAs in liver cancer and normal liver tissues. The difference in signal value of the enhancer that regulates miRNA was analyzed by ChIP-seq data of histone modification and transcription factor in liver cancer and normal liver tissues in ENCODE database. The differentially expressed miRNAs regulated by enhancers were screened out, and the correlation analysis was performed on the patient's survival and treatment targets. Results: 93 and 40 pairs of enhancer-miRNA were identified in liver cancer and normal liver tissues, respectively. ChIP-seqdata comparison analysis found that the signal of H3K27ac, H3K4me1 and sH3K4me3 histone modification in the region of enhancers regulating miRNA was significantly higher than that in the region of enhancers not regulating miRNA (|rho|>0.3, P<0.05). Moreover, the enrichment of multiple transcription factors in liver cancer-related enhancers was significantly lower than that in normal liver tissue-realted enhancers (|rho|>0.3, P<0.05). Differential expression analysis of enhancer-regulated miRNAs identified 6 miRNAs related to the survival of liver cancer patients(hsa-miR-4664, hsa-miR-5003,hsa-miR-1915,hsa-miR-3619,hsa-miR-4745, hsa-miR-6728),and found that these miRNAs were significantly associated with 87 genes for targeted therapy and 8 tumor immune checkpoint genes (|rho|>0.1, FDR<0.05). Conclusion: The enhancer-miRNA regulatory pairs and the characteristics of the enhancer that regulates miRNA were successfully identified in liver cancer. The miRNAs regulated by enhancers and related to the therapeutic targets and survival of patients with liver cancer were also screened out. It provides a valuable preliminary basis for future in-depth basic and clinical research in hepatic carcinoma.

miR-423-5p enhances chemoresistance of glioblastomas cells to temozolomide by regulating PDCD5 / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the miR-423-5p expression in brain glioma tissues and cell lines, and its promotive effect on temozolomide (TMZ) chemoresistance by targeting PDCD5 (programmed cell death protein 5). Methods: Tumor tissues and matched peritumoral tissues were collected from 20 brain glioma patients who were surgically treated in the Department of Neurosurgery, Affiliated Hospital of Beihua University between January 2017 and December 2018. Glioblastoma cell lines (U251, U87, SHG-44) and human normal glial cell line HMC-3 were also used in the study. The relative expression of miR-423-5p and PDCD5 in brain glioma and peritumoral tissues and cell lines was detected by qPCR. The synthesized miR-423-5p mimics and miR-NC were respectively transfected into U251 and U87 cells; meanwhile, TMZ at different concentrations (50, 100, 150 and 200 μmol/L) were also used to treat the cells. Then, the chemoresistance of cells to TMZ were determined. MTT assay and colony formation assay were used to examine the proliferation of U251 and U87 cells, andWestern blotting was used to detect the expression of c-caspase 3, Bcl-2 and PDCD5 proteins in U251 and U87 cells. The targeting relationship between PDCD5 and miR-423-5p was validated through Dual luciferase reporter gene assay. Results: miR-423-5p was highly expressed in glioma tissues and glioma cell lines (all P<0.01). As compared with the miR-NC group, the proliferation and TMZ-chemoresistance of U251 and U87 cells in miR-423-5p mimics group significantly increased (all P<0.01). Dual luciferase reporter gene assay validated that miR-423-5p could bind with PDCD5 3' UTR to suppress the expression of PDCD5. Conclusion: High expression of miR-423-5p enhances the chemoresistance of glioma cells to TMZ, and miR-423-5p may serve as a potential therapeutic target in the treatment of brain glioma.

miR-381 Attenuates Peripheral Neuropathic Phenotype Caused by Overexpression of PMP22

Charcot-Marie Tooth disease type 1A (CMT1A), the major type of CMT, is caused by duplication of peripheral myelin protein 22 (PMP22) gene whose overexpression causes structural and functional abnormalities in myelination. We investigated whether miRNA-mediated regulation of PMP22 expression could reduce the expression level of PMP22, thereby alleviating the demyelinating neuropathic phenotype of CMT1A. We found that several miRNAs were down-regulated in C22 mouse, a CMT1A mouse model. Among them, miR-381 could target 3′ untranslated region (3′UTR) of PMP22 in vitro based on Western botting and quantitative Real Time-PCR (qRT-PCR) results. In vivo efficacy of miR-381 was assessed by administration of LV-miR-381, an miR-381 expressing lentiviral vector, into the sciatic nerve of C22 mice by a single injection at postnatal day 6 (p6). Administration of LV-miR-381 reduced expression level of PMP22 along with elevated level of miR-381 in the sciatic nerve. Rotarod performance analysis revealed that locomotor coordination of LV-miR-381 administered C22 mice was significantly enhanced from 8 weeks post administration. Electrophysiologically, increased motor nerve conduction velocity was observed in treated mice. Histologically, toluidine blue staining and electron microscopy revealed that structural abnormalities of myelination were improved in sciatic nerves of LV-miR-381 treated mice. Therefore, delivery of miR-381 ameliorated the phenotype of peripheral neuropathy in CMT1A mouse model by down-regulating PMP22 expression. These data suggest that miRNA can be used as a potent therapeutic strategy to control diseases with copy number variations such as CMT1A.

Research Progress on MicroRNA Regulating Osteoblast Differentiation under Microgravity / 医用生物力学

MicroRNA (miRNA) is a kind of important gene expression regulatory molecules during biological process, but its regulation mechanism in metabolic process of bone tissues has not been completely clarified. In this review, the regulation of miRNA on osteoblast differentiation in microgravity environment was discussed. The positive and negative regulation of miRNA was summarized, respectively, with focus on introducing the mechanism of different genes. Some miRNA molecules that have important effects on bone metabolism under microgravity were enumerated. MiRNA plays an important role in regulating and controlling bone metabolic diseases in microgravity environment, and its related studies are significant for the prevention and treatment of bone loss induced by weightlessness.

Preliminary study on serum differential expression of microRNA and its target gene function in patients with severe Alzheimer disease / 中国神经精神疾病杂志

Objective To conduct an in-depth analysis on the correlation of miRNA and Alzheimer disease (AD) pathogenesis and provide an experimental basis for AD potential biomarkers by analysis of serum miRNA expression profiles in AD patients. Methods The miRNA expression profiles were exmined in 7 severe AD patients and 5 normal controls using high-throughput sequencing and bioinformatic analysis. Results Compared with the normal controls, there were serum differential expression of 112 miRNAs (DEmiRNAs) including 57 being up-regulated and 55 being down-regulated in patients with severe AD (P<0.05). GO-term function enrichment analysis showed that DEmiRNAs participated in the protein binding, ion binding, transcription metal binding, and biological metabolism and regulation process of organelle and cell membrane, etc. KEGG pathway enrichment analysis found that PI3K-Akt signal pathway was an important pathway of target genes. Conclusion The differential expression of serum miRNAs may be potential biomarkers of AD and the target genes of DEmiRNAs are related to the pathological changes of AD.

Study of miR-155 in the mechanism of rejection after liver transplantation in rats / 器官移植

Objective To explore the mechanism of microRNA (miRNA, miR)-155 in the rejection after liver transplantation in rats. Methods The rats were divided into two groups. In the xenograft model group (rejection group, n=10),the donors were male Lewis rats and the recipients were male BN rats.In the allograft model group(control group, n=10),both the donors and recipients were male Lewis rats.The rat models with orthotopic liver transplantation were established by two-cuff technique in two groups. At postoperative 7 d, the animals were sacrificed for the collection of blood and liver tissue samples. The serum levels of alanine aminotransferase (ALT), total bilirubin (TB), and cytokines of interleukin (IL)-2, IL-4, interferon (IFN)-γ were quantitatively measured. The pathological changes of liver tissues were observed under light microscope. In each group, three liver tissue samples were prepared and subject to high-throughput sequencing. The miRNAs related to rejection were identified for bioinformatics analysis to predict and analyze relevant signaling pathways and genes. Results In the rejection group, the serum levels of ALT and TB were significantly higher than those in the control group (both P<0.01). Compared with the control group, the levels of IL-2 and IFN-γ were considerably up-regulated (both P<0.01), whereas the level of IL-4 was dramatically down-regulated (P<0.01). Pathological examination demonstrated that more evident rejections were observed in the rejection group than the control group. High-throughput sequencing revealed that the expression level of miR-155 was significantly up-regulated in the rejection group, which was 5.89 times of that in the control group. Bioinformatics analysis demonstrated that up-regulation of miR-155 was associated with the mammalian target of rapamycin (mTOR), mitogen-activated protein kinase (MAPK) and T cell receptor signaling pathways. The genes which were probably responsible for regulation included the yeast autophagy related gene 1(ATG1) and its homologous gene ULK2, insulin-like growth factor-1 (Igf-1) and G protein-coupled receptor regulatory gene(Arrb1),etc.Conclusions miR-155 might promote the incidence and progression of rejection after liver transplantation in rats. The involved signaling pathways probably include the mTOR, MAPK signaling pathway and T cell receptor signaling pathway.ATG1,ULK2,Igf-1,and Arrb1 genes may participate in this process.

Expression of lncRNA LINC01001 in breast cancer and its effect on proliferation of MCF-7 cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the expression of long non-coding RNA LINC01001 in breast cancer tissues and its effect on the proliferation of breast cancer MCF-7 cells. Methods: The expression levels of lncRNA LINC01001 were analyzed in 12 cases of cancer and para-cancer tissues from breast cancer patients, who underwent surgical resection in Affiliated People’s Hospital of Hubei University of Medicine from March 2016 to June 2017. The plasmid over-expressing LINC01001 was transfected into human breast cancer MCF-7 cells. The cell cycle distribution and proliferation ability of MCF-7 cells were detected by flow cytometry and MTT assay, respectively. The mRNA expressions of miR-485-5p and CDKN1A mRNA were detected by qRT-PCR, and the protein expressions of CDKN1A, CDK4, CDK6 and Cyclin D1 were detected by Western blotting. Results: The expression level of lncRNA LINC01001 in breast cancer tissues was lower than that in para-cancer tissues (P<0.01). LINC01001 recombinant plasmid transfection significantly inhibited cell cycle progression (P<0.05) and cell proliferation (P<0.05) of MCF-7 cells. qRT-PCR showed that the expression level of miR-485-5p was decreased (P<0.01) and the expression level of CDKN1A mRNA was increased (P<0.01) after over-expressing LINC01001. Western blot results confirmed that over-expression of LINC01001 could promote the expression of CDKN1A protein, but decrease the expressions of CDK4, CDK6 and Cyclin D1 proteins. Conclusion: The expression of LINC01001 in breast cancer tissues was decreased. LINC01001 may down-regulate the expression of miR-485-5p to up-regulate the expression of CDKN1A, and further to inhibit the proliferation of breast cancer MCF-7 cells, providing experimental basis for the clinical application of lncRNA.

Research progress of the correlation between microRNAs and diabetic retinopathy / 中国糖尿病杂志

[Summary] MicroRNA (miRNA) is a class of endogenous small molecule non‐coding RNAs ,which can inhibit target gene expression via blocking protein translation or inducing mRNA degradation.MicroRNA is also involved in some important physiological processes ,such as cell development , differentiation ,proliferation and apoptosis. Recently ,studies have shown that miRNA plays a role in the development and progression of diabetic retinopathy (DR). This article summarized the research progress of correlation between miRNAs and DR.