Intervention Mechanism of Biejiajian Wan on Primary Liver Cancer by Regulating lncRNA SNHG5/miRNA-26a-5p/GSK-3β Signal Axis / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism of Biejiajian Wan in the intervention of primary liver cancer based on long non-coding RNA SNHG5 （lncRNA SNHG5）/micro RNA-26a-5p （miRNA-26a-5p）/glycogen synthase kinase-3β （GSK-3β） signal axis. MethodDouble luciferase reporting assay was used to verify the targeted interaction between lncRNA SNHG5 and miRNA-26a-5p， miRNA-26a-5p， and GSK-3β in HepG2 cells. Nude-mouse transplanted tumor model of human HepG2 were established and randomly divided into model group， Biejiajian Wan low-dose group （0.5 g·kg-1）， medium-dose group （1.0 g·kg-1）， and high-dose group （2.0 g·kg-1）， and sorafenib group （100 mg·kg-1）， with 10 mice in each group. The mice were given intragastric administration of normal saline or drug for 28 days， and the tumor volume was measured at different time. Hematoxylin-eosin （HE） staining was used to observe the histological changes of tumors. The nucleic acid levels of lncRNA SNHG5， miRNA-26a-5p， GSK-3β， and β-catenin mPNA in tumor tissue were detected by real-time quantitative polymerase chain reaction （Real-time PCR）. The protein expression levels of GSK-3β and β-catenin in tumor tissue were detected by western blot. ResultCompared with the SNHG5-WT （wild type） + miRNA NC （negative control） group， the relative luciferase activities of the SNHG5-WT + miRNA-26a-5p mimic group were decreased （P<0.05）. Compared with the GSK-3β-WT + miRNA NC group， the relative luciferase activity of the GSK-3β-WT + miRNA-26a-5p mimic group was decreased （P<0.05）. Compared with the model group， the tumor volume of Biejiajian Wan low-dose， medium-dose， and high-dose groups was significantly decreased （P<0.05， P<0.01）. Compared with the model group， the cells in the tumor tissue of nude mice in each dose group of Biejiajian Wan were sparsely arranged with necrocytosis， which showed concentration-dependent changes. Compared with the model group， the expression levels of lncRNA SNHG5， GSK-3β， and β-catenin were decreased （P<0.05， P<0.01）， while the expression of miRNA-26a-5p was increased in each dose group of Biejiajian Wan （P<0.05， P<0.01）. Compared with the model group， the protein expression levels of GSK-3β and β-catenin were decreased in each dose group of Biejiajian Wan （P<0.05， P<0.01）. ConclusionBiejiajian Wan may affect the necrosis of liver cancer cells through lncRNA SNHG5/miRNA-26a-5p/GSK-3β signal axis and thus play an anti-tumor role. This research will provide more theoretical basis for the clinical application of Biejiajian Wan.

Construction of competitive endogenous RNA network mediated by lung ischemia-reperfusion core genes / 器官移植

Objective To analyze the core genes of lung ischemia-reperfusion injury and construct a competitive endogenous RNA (ceRNA) network. Methods Original data of GSE145989 were downloaded from the Gene Expression Omnibus (GEO) database as the training set, and the GSE172222 and GSE9634 datasets were used as the validation sets, and the differentially-expressed genes (DEG) were identified. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed. Protein-protein interaction (PPI) network was constructed, and the core genes were screened, and the diagnostic values of these core genes and the immune infiltration levels of immune cells were evaluated. The ceRNA network was constructed and validated. The targeted drugs based on ceRNA network were assessed. Results A total of 179 DEG were identified, including 61 down-regulated and 118 up-regulated genes. GO analysis showed that DEGs were associated with multiple biological processes, such as cell migration, differentiation and regulation, etc. They were correlated with cell components, such as vesicle membrane, serosa and membrane raft, etc. They were also associated with multiple molecular functions, such as chemokine receptor, G protein-coupled receptor, immune receptor activity and antigen binding, etc. KEGG pathway enrichment analysis revealed that DEG were involved in tumor necrosis factor (TNF), Wnt, interleukin (IL)-17 and nuclear factor (NF)-κB signaling pathways, etc. PPI network suggested that CD8A, IL2RG, STAT1, CD3G and SYK were the core genes of lung ischemia-reperfusion injury. The ceRNA network prompted that miR-146a-3p, miR-28-5p and miR-593-3p were related to the expression level of CD3G. The miR-149-3p, miR-342-5p, miR-873-5p and miR-491-5p were correlated with the expression level of IL-2RG. The miR-194-3p, miR-512-3p, miR-377-3p and miR-590-3p were associated with the expression level of SYK. The miR-590-3p and miR-875-3p were related to the expression level of CD8A. The miR-143-5p, miR-1231, miR-590-3p and miR-875-3p were associated with the expression level of STAT1. There were 13 targeted drugs for CD3G, 4 targeted drugs for IL-2RG, 28 targeted drugs for SYK and 3 targeted drugs for lncRNA MUC2. No targeted drugs were identified for CD8A, STAT1 and other ceRNA network genes. Conclusions CD8A, IL2RG, STAT1, CD3G and SYK are the core genes of lung ischemia-reperfusion injury. The research and analysis of these core genes probably contribute to the diagnosis of lung ischemia-reperfusion injury and providing novel research ideas and therapeutic targets.

Impact of lncRNA FGD5-AS1 on the malignant biological behavior of bladder cancer cells by regulating miR-129-5p/CDK6 axis / 现代泌尿外科杂志

【Objective】 To investigate the impact of long non-coding RNA (lncRNA) FGD5-AS1 on the malignant biolo-goical behavior of bladder cancer (BC) cells by regulating micro RNA (miR)-129-5p/cyclin dependent kinase 6 (CDK6) axis. 【Methods】 Human BC cell line T24 was cultured from tumor tissue and paracancerous tissue of 105 patients with confirmed BC. The expressions of FGD5-AS1, miR-129-5p and CDK6 mRNA in tissue samples and T24 cells were detected with RT-qPCR. T24 cells were randomly divided into control group, si-NC group, si-FGD5-AS1 group, si-FGD5-AS1+inhibitor NC group and si-FGD5-AS1+miR-129-5p inhibitor group. The cell viability, migration, invasion andapoptosis were detected with CCK-8, Wound healing test, Transwell assay and flow cytometry, respectively. The expressions of Bax, Bcl-2, Caspase3 and CDK6 were detected with Western blot. The relationship between FGD5-AS1 and miR-129-5p, between miR-129-5p and CDK6 were verified with double luciferase reporter gene experiment. 【Results】 FGD5-AS1 and CDK6 mRNA were highly expressed in BC tissue, while miR-129-5p was lowly expressed (P<0.05). After FGD5-AS1 silencing, the expression of FGD5-AS1,A450 value, cell scratch healing rate, cell invasion number, and expressions of Bcl-2 and CDK6 were significantly lower, while the apoptosis rate and expressions of miR-129-5p, Bax and Caspase3 were significantly higher (P<0.05). Inhibition of miR-129-5p expression reversed the effects of FGD5-AS1 silencing on various indexes of BC cells (P<0.05). FGD5-AS1 negatively regulated the expression of miR-129-5p, and miR-129-5p negatively regulated the expression of CDK6. 【Conclusion】 Silencing FGD5-AS1 may inhibit the expression of CDK6 protein by up-regulating miR-129-5p, thus inhibiting the proliferation, migration and invasion of BC cells and promoting cell apoptosis.

circSNRK alleviates ischemia-reperfusion injury in renal tubular epithelial cell by up-regulating Akt pathway / 器官移植

Objective To investigate the role and mechanism of circular RNA SNRK (circSNRK) in ischemia-reperfusion injury (IRI). Methods A hypoxia-reoxygenation (IRI) cell model was established. The expression level of circSNRK after IRI treatment and the effect of overexpression of circSNRK on cell proliferation and apoptosis were detected. The targets of circSNRK were identified. HK2 cells were divided into the blank group (Mock group), IRI group, control plasmid+IRI group (IRI+NC group), human circSNRK overexpression+IRI group (IRI+circSNRK group), human circSNRK overexpression+IRI+protein kinase B (Akt) inhibitor group (IRI+circSNRK+MK2206 group) and control plasmid group (NC group). Cell proliferation and apoptosis were detected in the Mock, IRI, IRI+NC and IRI+circSNRK groups. The gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of the target of circSNRK were carried out. The expression levels of CDKN1A, Akt, B-cell lymphoma (Bcl)-2, cysteinyl aspartate specific proteinase (Caspase)-9 messenger RNA (mRNA), and those of p21, Bcl-2, Caspase-9, Akt and p-Akt proteins were detected in the Mock, IRI, IRI+NC and IRI+circSNRK groups, respectively. Cell proliferation and apoptosis were determined in the NC, IRI+NC, IRI+circSNRK and IRI+circSNRK+MK2206 groups. Results Compared with the Mock group, the expression level of circSNRK was lower, and cell proliferation capability of HK2 cells was decreased and cell apoptosis was increased in the IRI group. In the IRI+circSNRK group, cell proliferation capability was higher, whereas cell apoptosis was lower than those in the IRI+NC group. circSNRK could act on 648 targets through 51 microRNAs (miRNAs). GO enrichment analysis revealed that the targets of circSNRK were mainly enriched in biological processes (such as cell process and biological regulation), cell components (such as cell parts, cells and extracellular parts), and molecular functions (such as binding, binding proteins and enzymes). KEGG enrichment analysis showed that the targets of circSNRK were mainly enriched in cancer signaling pathway, phosphatidylinositol 3-kinase (PI3K)-Akt signaling pathway, miRNA in cancer and other related signaling pathways. Compared with the Mock group, the relative expression levels of CDKN1A and Caspase-9 mRNA were higher, the expression level of miR-99a-5p RNA was higher and the relative expression levels of Akt and Bcl-2 mRNA were lower in the IRI group. Compared with the IRI+NC group, the relative expression levels of CDKN1A and Caspase-9 mRNA were lower, those of Akt and Bcl-2 mRNA were higher, and the expression level of miR-99a-5p RNA was lower in the IRI+circSNRK group, and the differences were statistically significant (all P < 0.05). Compared with the Mock group, the expression levels of p21 and Caspase-9 proteins were higher, while those of p-Akt, Akt and Bcl-2 proteins were lower in the IRI group. Compared with the IRI+NC group, the expression levels of p21 and Caspase-9 proteins were lower, whereas those of p-Akt, Akt and Bcl-2 proteins were higher in the IRI+circSNRK group. The miR-99a-5p binding sites were observed in circSNRK and Akt. Compared with the NC group, cell proliferation capability was declined in the IRI+NC group. Compared with the IRI+NC group, cell proliferation capability was elevated in the IRI+circSNRK group. Compared with the IRI+circSNRK group, cell proliferation capability was declined in the IRI+circSNRK+MK2206 group (all P < 0.05). The cell apoptosis level in the IRI+NC group was higher than that in the NC group. The cell apoptosis level in the IRI+circSNRK group was lower compared with that in the IRI+NC group. The cell apoptosis level in the IRI+circSNRK+MK2206 group was higher than that in the IRI+circSNRK group. Conclusions Under IRI conditions, circSNRK may affect the proliferation and apoptosis of HK2 cells probably via the Akt signaling pathway.

Recent Progress on Pharmaceutical Properties of Extracellular Vesicles from Mesenchymal Stem Cells--Review / 中国实验血液学杂志

Mesenchymal stem cells (MSCs) have been officially approved in many countries to treat graft-versus-host disease, autoimmune disorders and those associated with tissue regeneration after hematopoietic stem cell transplantation. Studies in recent years have confirmed that MSC acts mainly through paracrine mechanism, in which extracellular vesicles secreted by MSC (MSC-EV) play a central role. MSC-EV has overwhelming advantages over MSC itself in the setting of adverse effects in clinical application, indicating that MSC-EV might take the place of its parent cells to be a potentially therapeutic tool for "cell-free therapy". The pharmaceutical properties of MSC-EV largely depend upon the practical and optimal techniques including large-scale expansion of MSC, the modification of MSC based on the indications and the in vivo dynamic features of MSC-EV, and the methods for preparing and harvesting large amounts of MSC-EV. The recent progresses on the issues above will be briefly reviewed.

Salvianolic acid A contributes to cartilage endplate cell restoration by regulating miR-940 and miR-576-5p / 中国骨伤

OBJECTIVE@#To investigate whether Salvianolic acid A (SAA) can restore cartilage endplate cell degeneration of intervertebral discs and to identify the mechanism via regulation of micro-RNA.@*METHODS@#Cartilage endplate cells were isolated from lumbar intervertebral disc surgical samples and were treated with serum containing a series of concentrations of SAA (2, 5, and 10 ?M) for 24, 48, and 72 h to identify a proper dose and treatment time of SAA. The effect SAA on interlenkin-1β (IL-1β)-induced extracellular matrix degradation of cartilage endplate cells were analyzed by Alcian blue staining and assessment of the expression levels of ADAMTS-5, MMP3 and Col2a1. Further, the potential target miRNAs were preliminarily screened by micro-RNA sequencing combining qRT-PCR and Western blot, and then, the miRNAs mimics and inhibitors were used to verify the regulatory effect of SAA on potential target miRNAs.@*RESULTS@#The 10 μM SAA treatment for 48 h significantly enhanced the viability of cartilage endplate cells, and increased Col2a1 expression and glycosaminoglycan accumulation that were repressed by IL-1β, and reduced the effect of IL-1β on ADAMTS-5, and MMP3. Screening analysis based on micro-RNA sequencing and Venny analysis identified the downstream micro-RNAs, including miR-940 and miR-576-5p. Then, the miR-940-mimic or miR-576-5p-mimic were transfected into CEPCs. Compared with the SAA group, the expression of ADAMTS-5 and MMP3 increased significantly and the expression of COL2A1 obviously decreased after overexpression of miR-940 or miR-576-5p in CEPCs.@*CONCLUSION@#Salvianolic acid A attenuated the IL-1β-induced extracellular matrix degradation of cartilage endplate cells by targeting regulate the miR-940 and the miR-576-5p.

Mechanism of lncRNA-miRNA-related Signaling Pathways in Gastric Cancer and Chinese Medicine Intervention： A Review / 中国实验方剂学杂志

Gastric cancer （GC） is one of the most common malignant tumors in the digestive system， with high morbidity and mortality. Early clinical symptoms of GC are not obvious， and most of them have entered the advanced stage after discovery， which greatly reduces the clinical cure rate and affects the quality of life of patients， and the prognosis is very poor. In recent years， with the continuous exploration in the field of bioinformatics， it has been found that micro-RNA （miRNA） and long non-coding RNA （lncRNA） exist as non-coding RNA （ncRNA） without translation ability， and regulate the expression levels of related signal proteins by acting on a certain target， thereby activating or inhibiting a certain signaling pathway， which plays an important role in assisting diagnosis， guiding clinical medication， and judging prognosis in the progress of GC. Chinese medicine is easily accepted by patients because of its good curative effect and less side effects. In the present basic studies， with the interaction mechanism between miRNA， lncRNA and signaling pathways as the breakthrough point， various studies on the regulation of related signaling molecules and signaling pathways by Chinese medicine have been carried out. A large number of experimental data have proved that the development of GC is closely related to the interaction of miRNA， lncRNA， and related signaling pathways， and Chinese medicine， with multi-target， multi-mechanism， and multi-pathway characteristics， affects various signaling molecules and signaling pathways and intervenes in the progress of GC cells. This paper reviewed the basic research on lncRNA， miRNA molecules， and main signaling pathways involved in the occurrence and development of GC， and summarized specific molecular mechanisms of Chinese medicine in the regulation of each signaling pathway， hoping to provide references for modern research of Chinese medicine in the intervention of GC progress at the molecular level.

The Regulation of mTOR Signaling Pathway on Adipogenesis / 中国生物化学与分子生物学报

Adipose tissue is a major energy storage and endocrine organ. Adipogenesis is a complex process of cell differentiation, which is regulated by nutrient levels, hormones and metabolites, etc. The mammalian target of rapamycin (mTOR) complex includes two protein complexes, mammalian target of rapamycin complex 1 (mTORC1) and mTORC2. The lipid kinase-like domain contained in the mTOR complex lays the foundation for the mTOR pathway to regulate adipogenesis. Research on some components of mTORC1 and mTORC2 has verified the roles of mTOR in the regulation of adipogenesis. Based on previous studies, we reviewed the research of miR-199a-3p, miR-103, miR-188, Src-associated substrate in mitosis of 68 kD (Sam68), endostatin and other substances in the regulation of adipogenesis through mTORC1 and mTORC2. At the same time, we had further constructed the adipogenesis network regulated by mTOR signaling pathway, including insulin/IGF pathway, PI3K-AKT pathway, amino acid pathway, AMPK pathway, cAMP pathway, cGMP pathway, NOTCH pathway, and the modulation of bta-miR-150, 4-O-methylasochlorin and a variety of proteins. This article mainly reviewed the characteristics of mTOR complex and the latest research progress in the regulation of adipogenesis by mTOR pathway. It was pointed out that mTORC2 can regulate lipid uptake, lipolysis and regulate the function of mTORC1. However, there are fewer studies on mTORC2 compared to mTORC1, so further researches on adipogenesis and lipid metabolism may be more focused on mTORC2.

Protective effects of micro RNA-98-5p targeting Kruppel-like factor 9 against myocardial ischemia-reperfusion injury in rats / 解剖学报

Objective To investigate the protective effect of micro RNA(miR)-98-5p targeting Kruppel-like factor 9(KLF9) against myocardial isehemia-reperfusion(MI/R) injury in rats. Methods Totally 50 rats were randomly divided into sham operation group, model group, miR-98-5p agomir group, agomir-NC group, and miR-98-5p agomir+pcDNA-3. 1-KLF9 group, 10 in each group. MI/R model was established by coronary artery ligation. The pathological changes of myocardial tissue were detected by HE staining. The myocardial apoptosis were detected by TUNEL. The levels of creatine kinase (CK), creatine kinase isoenzymes (CK-MB) and lactate dehydrogenase (LDH) in serum were detected by ELISA. The expression levels of miR-98-5p and KLF9 mRNA were detected by Real-time PCR. The expression of KLF9, Bax and JAK2/STAT3 pathway relative protein were detected by Western blotting. Dual luciferase assay verified the relationship between miR-98-5p and KLF9. Results Compared with the sham operation group, the arrangement of myocardial cells in the model group was disordered, and the myocardial cells appeared necrosis. The apoptosis rate of myocardial cells, serum CK, CK-MB and LDH contents increased, the expression level of miR-98-5p decreased, and the expression levels of KLF9 mRNA and protein, p-JAK2 and p-STAT3 protein increased (P < 0. 05) . After the overexpression of miR-98-5p, the myocardial cells arranged more orderly and the myocardial cell necrosis decreased. The apoptosis rate of myocardial tissue, the contents of CK, CK-MB and LDH in serum and the expression of p-JAK2 and p-STAT3 protein were decreased (P< 0. 05) . The result of dual luciferase assay showed that KLF9 was the target gene of miR-98-5p. The overexpression of KLF9 reversed the effects of miR-98-5p agomir on myocardial injury. Conclusion MiR-98-5p targeting KLF9 can improve the myocardial injury of MI/R rats. The mechanism may be related to the regulation of JAK2/STAT3 signal pathway by miR-98-5p which inhibit myocardial cell apoptosis.

The molecular mechanism of pathological changes in osteoarthritis / 中华物理医学与康复杂志

Objective:To explore the molecular mechanism of pathological changes in osteoarthritis (OA) through applying bioinformatics to analyze the miRNA-mRNA regulatory network.Methods:MiRNA expression data from human serum samples were downloaded from the Gene Expression Omnibus database. Differentially-expressed miRNA was identified using the linear modelling package of the Bioconductor software suite. The target differentially-expressed mRNA was predicted using version 2.0 of the miRWalk database. Version 3.7.1 of the Cytoscape software was used to construct the miRNA-mRNA regulatory network for OA. The target genes were analyzed by using gene ontology and the Kyoto Encyclopedia of Genes and Genomes. The protein-protein interaction network was constructed and the core genes in osteoarthritis pathology were screened out.Results:A total of 7 differentially-expressed miRNAs (all down-regulated) and 900 mRNAs were identified, mostly involved in the negative regulation of protein binding, DNA binding, or transcription in the cell cycle. Ten core genes were screened out: MAPK1, TP53, MAPK14, CCND1, EP300, POLR2E, POLR2F, ABL1, RAC1 and SKIV2L2.Conclusions:Multiple miRNAs, target genes and signaling pathways are involved in the development of OA. The miRNA-mRNA regulatory network identified provides new ideas for exploring the molecular mechanism of OA′s pathology and its clinical diagnosis and treatment.

Expression of miR-155 and IFN-γ in lung injury model of neonatal rats with acute respiratory distress syndrome / 国际儿科学杂志

Objective:To study the expression of micro RNA-155(miR-155) and IFN-γ in lung tissue in a neonatal rat model of acute respiratory distress syndrome(ARDS)lung injury by intraperitoneal injection of lipopolysaccharide(LPS).Methods:Eighty neonatal SD rats on the 7th day after birth were assigned to the experimental group(LPS group)and control group(isotonic NaCl group), with 40 rats in each group.LPS solution(4 mg/kg)was injected into the abdominal cavity of neonatal SD rats in the experimental group to establish an animal model of neonatal acute respiratory distress syndrome(NARDS). The control group was established by isotonic NaCl solution(4 ml/kg)in the same way.The lung tissue samples were taken at 3 h, 6 h, 12 h and 24 h after drug administration to observe the surface changes.Then the lung sections were stained with HE to observe the pathological changes and score the lung tissue injury.Finally, the expression levels of miR-155 and IFN-γ in the lung tissue were tested by RT-PCR and ELISA techniques, respectively.Results:(1)At the beginning of the experiment, the neonatal rats in the experimental group gradually showed the clinical manifestations of ARDS, and the macroscopic observation, pathological changes and lung tissue injury scores of the lung tissues suggested the appearance of NARDS lung injury, indicating that the model was successful.(2)The expression levels of miR-155(1.33±0.12 vs 0.95±0.02、1.77±0.17 vs 0.96±0.01、2.18±0.09 vs 0.96±0.02 and 2.43±0.06 vs 0.96±0.02)and IFN-γ(370.79±13.89 vs 273.03±11.44、424.24±10.11vs270.70±13.05、466.63±6.57 vs 268.11±7.88 and 519.13±7.09 vs 272.97±12.54)ng/L in the lung tissue of rats between the experimental group and the control group were significantly different( P<0.01), and the difference was statistically significant among the groups in the experimental group( F values were 165.983 and 408.574, P<0.01). The expression levels of miR-155 and IFN-γ in the lung tissue of the experimental group increased gradually over time and showed an increasing trend. Conclusion:After the successful establishment of NARDS animal model, the expression levels of miR-155 and IFN-γ in the lung tissue of NARDS rats have significantly increased and showed a sequential pattern.MiR-155 is expected to become an early biomarker for the diagnosis of NARDS.

Progress on biomarkers of EB virus-related posttransplant lymphoproliferative disease / 器官移植

Posttransplant lymphoproliferative disease (PTLD) is a series of heterogeneous lymphoproliferative diseases and a severe complication after solid organ transplantation in children. Over 70% of PTLD is associated with Epstein-Barr virus (EBV). EBV-related B-cell lymphoma is also the main malignant tumor after pediatric organ transplantation. EBV-related PTLD is still a challenge in pediatric solid organ transplantation, which is mainly caused by immune function damage induced by immune suppression after transplantation. However, the specific mechanism remains elusive. In recent years, biomarkers have been developed to guide the diagnosis and individualized treatment of EBV-related PTLD, which possesses excellent application prospect. In this article, research progresses on the incidence of EBV-related PTLD in solid organ transplantation and its biomarkers were reviewed, aiming to explore novel ideas for clinical diagnosis and treatment.

Effects of Competing Endogenous RNA on Proliferation and Differentiation of Skeletal Muscle Cells / 中国生物化学与分子生物学报

The skeletal muscle is an important part of the animal body, which is closely related to body movement, energy consumption, production performance and its development process is regulated by many factors. The molecular mechanism of skeletal muscle growth and development is not only closely related to animal husbandry production, but also provides a theoretical basis for the treatment of muscle dis-eases, such as amyotrophic muscular dystrophy, and so on. Non-coding RNA (ncRNA) is a kind of RNA without coding ability, in which circular RNA (circRNA), long non-coding RNA (lncRNA) and micro RNA (miRNA) have been proved to play an important role in the development of skeletal muscles. miR-NA can specifically bind to the 3′ untranslated region (3′ UTR) of the target gene through its seed se-quence, so as to inhibit the translation process of the target gene or directly degrade the target gene, and become a prominent participant in a variety of biological processes. Studies have shown that circRNA, ln-cRNA, pseudogenes and mRNA with miRNA response elements (MRE) can competitively combine miRNA to regulate gene expression, forming a competing endogenous RNA (ceRNA) regulatory network model. This hypothesis subverts the previous concept of unidirectional regulation of target genes by miRNA, and endows them with new biological functions in transcriptome, which is of great biological significance. In recent years, it has been found that ceRNA plays an important regulatory role in the growth and development of skeletal muscles. This paper reviews the role of miRNA, lncRNA and circRNA in the proliferation and differentiation of animal skeletal muscle cells, which is an important member of ceRNA mechanism, in order to broaden the research direction of skeletal muscle mechanism and provide new ideas for the development of animal husbandry and the treatment of muscle diseases.

Expression and mechanism of microRNA-27a in cervical cancer / 解剖学报

Objective To investigate the effects of microRNA (miR)-27a on proliferation, apoptosis and invasion of cervical cancer cells by targeting F-box and WD repeat domain containing protein 7 (FBXW7) expression. Methods Thirty cases of cervical cancer and paracancerous tissues and 30 cases of normal tissues were used in the experiment. The expression of miR-27a in cervical cancer, paracancerous tissue, normal cervical tissue, cervical cancer cells (SiHa, Caski, HeLa, HCC94) and cervical squamous epithelial immortalized cell H8 were detected by Real-time PCR. MiR-27a inhibitor and its negative control were transfected into SiHa cells by liposome transfection. CCK-8 assay, flow cytometry and Transwell assay were used to detect the effects of miR-27a on the proliferation, cell cycle, apoptotic rate and invasive ability of SiHa cells. Bioinformatics was used to predict the targeting gene of miR-27a. Double luciferase reporter gene assay combined with Western blotting was used to verify the targeting regulation of miR-27a on FBXW7. Results Compared with the normal cervical tissues and the adjacent tissues, the expression of miR-27a was higher in cervical cancer tissues (P<0.05) ; Compared with the cervical squamous epithelial immortalized cells H8, the expression of miR-27a in cervical cancer cells SiHa, Caski, HeLa and HCC94 was higher (P<0.05). Inhibiting the expression of miR-27a in SiHa cells could significantly reduce the proliferation activity of cells (P<0.05), increase the proportion of G

MicroRNA-98-5p targeting ribonucleotide reductase small subunit M2 regulating cisplatin resistance in cervical cancer cells / 解剖学报

Objective To investigate the regulation and mechanism of microRNA (miR)-98-5p on cisplatin sensitivity in cisplatin-resistant cervical cancer cells. Methods The cisplatin(DDP) +miR-NC group (transfected miR- NC), DDP + miR-98-5p group (transfected miR-98-5p mimics), DDP + si-NC group (transfected si-NC), DDP + si- ribonucleotide reductase subunit M2 (RRM2) group (transfected si-RRM2), DDP + miR-98-5p + pcDNA group (co- transfected miR-98-5p mimics and pcDNA), DDP + miR-98-5p + pcDNA-RRM2 group (co-transfected miR-98-5p mimics and pcDNA-RRM2) were transfected into HeLa/DDP cells by liposome method. Real-tim PCR, Western blotting, CCK-8, Transwell chamber and dual luciferase reports gene detection assay were used to detect the expression of miR-98-5p, RRM2, cyclin Dl, P21, matrix metalloproteinase (MMP)-2 and MMP-9 in cells, inhibition rate, half inhibitory concentration(IC

TRPM3/miR-204 in the regulation of ophthalmic diseases / 国际眼科杂志(Guoji Yanke Zazhi)

@#MicroRNAs(miRNAs)are one of the most important regulatory factors of gene expression, which involved in the growth, development, differentiation and apoptosis of various cells, tissues and organs. TRPM3 is located in human chromosome 9 and belongs to M sub-family of the transient receptor potential(TRP)channels. MiR-204 is located on TRPM3 intron 6 and participates in the regulation of post-transcriptional gene expression through cleavage or translation inhibition of target mRNAs. Studies have shown that TRPM3/miR-204 complex locus plays an important role in the occurrence and development of eye diseases such as cataract, glaucoma, corneal neovascularization, corneal wound healing, retinal diseases, optic nerve diseases and so on. In this paper, the biological function of TRPM3/miR-204, its expression and regulation in the eyes and its correlation with a variety of ophthalmic diseases are reviewed.

Effect of micro RNA-296-3p regulated cell on proliferation, migration and invasion of tongue squamous cell carcinoma by targeting nucleoprotein 1 / 中国医师进修杂志

Objective:To investigate the effects of micro RNA(miR)-296-3p on cell proliferation, migration and invasion of tongue squamous cell carcinoma and its underlying mechanism.Methods:Normal oral epithelial mucosal keratinocytes HOK was used as the control. Real-time quantitative polymerase chain reaction (PCR) was used to detect the expression of miR-296-3p and nucleoprotein 1(NUPR1) mRNA in tongue squamous cell carcinoma cell lines CAL27, SCC15 and SCC9, and Western blot was used to detect the expression of NUPR1 protein. CAL27 cells were divided into NC group, miR-con group, miR-296-3p group, si-con group, si-NUPR1 group, miR-296-3p+pcDNA group and miR-296-3p+pcDNA-NUPR1 group, then CCK8 method was used to detect the cell activity. Transwell was used to detect the cell migration and invasion. Western blot was used to detect the protein expression levels of proliferating cell nuclear antigen (PCNA), matrixmetallo proteinase-2(MMP-2) and matrixmetallo proteinase-9(MMP-9). Dual-luciferase reporter assay system was implemented to verify the relationship between miR-296-3pand NUPR1.Results:Compared with the HOK cells, the content of miR-296-3p in the tongue squamous cell line CAL27, SCC15 and SCC9 groups was significantly reduced (0.54 ± 0.08, 0.38 ± 0.05, 0.59 ± 0.07 vs. 1.04 ± 0.12, t = 10.401, 15.231 , 9.718, P<0.05), while the expression levels of NUPR1mRNA (5.94 ± 0.40, 4.48 ± 0.45, 5.19 ± 0.48 vs.0.94 ± 0.12, t = 35.918, 22.803, 25.769) and protein (0.79 ± 0.09, 0.54 ± 0.05, 0.62 ± 0.08 vs. 0.28 ± 0.04, t = 15.535, 12.182, 11.404) were significantly increased (all P<0.05). Compared with those in the NC group or miR-con group, CAL27 cell activity, migration number, invasion number and the protein expression of PCNA, MMP-2 and MMP-9 in the miR-296-3p group were decreased (all P<0.05). Compared with those of the NC group or the si-con group, the CAL27 cell activity, migration number, number of invasions and the protein expression of PCNA, MMP-2 and MMP-9 in the si-NUPR1 group were decreased (all P<0.05). miR-296-3p negatively regulatedthe expression of NUPR1 in CAL27 cells. Compared with the miR-296-3p+pcDNA group, CAL27 cell viability (138.34 ± 5.73 vs. 98.54 ± 5.89, t = 14.530), migration number (95.28 ± 7.56 vs. 67.92 ± 5.23, t = 8.929), invasion number (184.53 ± 17.57 to 101.26 ± 10.64, t = 12.162) and the protein expression of PCNA (0.68 ± 0.07 to 0.35 ± 0.06, t = 10.738), MMP-2 (0.43 ± 0.05 to 0.29 ± 0.04, t = 6.559) and MMP-9 (0.58 ± 0.06 vs. 0.33 ± 0.08, t = 7.500) in the miR-296-3p+pcDNA-NUPR1 group were increased (all P<0.05). Conclusions:miR-296-3p may inhibitthe proliferation, migration and invasion of tongue squamous cell carcinoma cells by negatively regulating NUPR1.

Clinical value of plasma miR-27 expression in patients with diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the clinical value of microRNA(miRNA, miR)-27 expression in patients with diabetic retinopathy(DR).<p>METHODS: A total of DR 80 patients(DR group)treated between January 2019 and January 2020 were retrospectively reviewed. Meanwhile, 40 patients with simple type 2 diabetes mellitus(T2DM)(T2DM group)and 40 normal healthy persons(control group)were enrolled, and plasma RNA was extracted. Real time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR)was adopted to determine plasma miR-27 expression, and enzyme-linked immunosorbent assay was performed to determine the vascular endothelial growth factor(VEGF)level. Plasma miR-27 and serum VEGF expression in different groups and in patients with different severities of DR was comparatively analyzed. Multivariate Logistic regression analysis was performed to screen factors influencing the expression of miR-27 in patients with DR, and Pearson correlation analysis of miR-27, serum VEGF and blood glucose indexes was conducted. Meanwhile, significance of miR-27 in pathogenesis of DR was summarized.<p>RESULTS: DR group had the highest plasma miR-27 and serum VEGF levels, followed by T2DM group, and then the control group(<i>P</i><0.05). Proliferative diabetic retinopathy(PDR)patients had higher levels of plasma miR-27, serum VEGF, fasting blood glucose and glycated hemoglobin than those with non-proliferative diabetic retinopathy(NPDR)(<i>P</i><0.05). It was found that course of disease(<i>OR</i>=3.206), fasting blood glucose(<i>OR</i>=2.570), glycated hemoglobin(<i>OR</i>=2.787), VEGF(<i>OR</i>=3.442)and severity of DR(<i>OR</i>=5.842)were influencing factors of plasma miR-27 expression in DR patients(<i>P</i><0.05). In DR patients, relative expression of plasma miR-27 was positively correlated with serum VEGF, fasting blood glucose and glycated hemoglobin(<i>r</i>=0.548, 0.398, 0.522, all <i>P</i><0.05).<p>CONCLUSION: DR patients have higher plasma miR-27 expression level than those with simple T2DM and normal healthy people. The duration of diabetes, fasting blood glucose, glycated hemoglobin and severity of DR all affect the expression of miR-27. Besides, miR-27 is positively correlated with serum VEGF, glycated hemoglobin and fasting blood glucose. It is speculated that miR-27 may mediate the pathogenesis and progression of DR by regulating glucose metabolism and promoting angiogenesis.

Effect of micro RNA-335-5p regulating bone morphogenetic protein 2 on osteogenic differentiation of human bone marrow mesenchymal stem cells / 中国修复重建外科杂志

Objective: To investigate the effect of micro RNA (miR)-335-5p regulating bone morphogenetic protein 2 (BMP-2) on the osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs). Methods: hBMSCs were cultured in vitro and randomly divided into control group (group A), miR-335-5p mimics group (group B), miR-335-5p mimics negative control group (group C), miR-335-5p inhibitor group (group D), and miR-335-5p inhibitor negative control group (group E). After grouping treatment and induction of osteogenic differentiation, the osteogenic differentiation of cells in each group was detected by alkaline phosphatase (ALP) and alizarin red staining; the expressions of miR-335-5p and BMP-2, Runt-related transcription factor 2 (Runx2), osteopontin (OPN), and osteocalcin (OCN) mRNAs were detected by real-time fluorescence quantitative PCR analysis; the expressions of Runx2, OPN, OCN, and BMP-2 proteins were detected by Western blot. Results: Compared with group A, the relative proportion of ALP positive cells and the relative content of mineralized nodules, the relative expressions of BMP-2, miR-335-5p, OPN, OCN, Runx2 mRNAs, the relative expressions of Runx2, OPN, OCN, and BMP-2 proteins in group B were significantly increased ( P0.05). Conclusion: miR-335-5p can up-regulate BMP-2 expression and promote osteogenic differentiation of hBMSCs.

Regulatory effect of miR-125b on ventricular remodeling after myocardial infarction in rats / 吉林大学学报(医学版)

Objective: To investigate the effect of miR-125b on the ventricular remodeling after myocardial infarction in the rats via phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway, and to elucidate its mechanism. Methods: A total of 75 rats were divided into sham operation group, myocardial infarction group and miR-125b inhibitor group, with 25 rats in each group. The acute myocardial infarction models were established in the rats in myocardial infarction group and miR-125b inhibitor group. The rats in miR-125b inhibitor group was injected with miR-125b inhibitor via the tail vein. The rats were sacrificed 4 weeks later, the body weig hts were recorded; the hearts were obtained, and the big vessel and left and right auricular appendix were cut out; the heart weights were weighed. The heart weight/body weight (HW/BW), (left ventricle + ventricular septum) weight/body weight (LV+S)/BW, (left ventricular + ventricular septum) weight/heart weight (LV+S)/HW were calculated. HE staining was used to observe the morphological changes of myocardium tissue. Masson staining was used to observe the myocardial fibrosis, and the collagen volume integral of myocardium was calculated. Immunofluorescence staining was used to observe the expression levels of type I collagen and type III collagen proteins in the myocardium tissue of the rats. Western blotting method was used to detect the expression levels of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), PI3K, Akt and phosphorylated Akt (p-Akt) in the peripheral region of myocardial infarction of the rats. Results: The HE staining results showed that the myocardial cells in sham operation group were arranged neatly; the myocardial cells in myocardial infaction group had hypertrophy and disordered arrangement; the myocardial cells in miR-125b inhibitor group were arranged neatly. Compared with sham operation group, the HW/BW, (LV + S)/BW and (LV + S)/HW of the rats in myocardial infarction group were elevated (P<0.05), the collagen volume integral was increased (P<0.05), the protein expression levels of type I collagen and type III collagen were increased (P<0.05), the expression levels ANP and BNP proteins were increased (P<0.05), and the expression levels of PI3K and p-Akt proteins were decreased (P<0.05). Compared with myocardial infarction group, the HW/BW, (LV+S)/BW and (LV+S)/HW of the rats in miR-125 b inhibitor group were decreased (P<0.05), the collagen volume integral was decreased (P<0.05), the expression levels of type I collagen and type III collagen proteins were decreased (P<0.05), the expression levels of ANP and BNP proteins were decreased (P<0.05), and the expression levels of PI3K and p-Akt proteins were increased (P<0.05). Conclusion: Inhibition of miR-125b can inhibit ventricular remodeling after myocardial infarction in the rats by inhibiting the PI3K/Akt signaling pathway, and plays an important role in reversing ventricular remodeling after myocardial infarction.