Objective To construct vectors that express phosphatidylinositol-3-kinase, catalytic, beta polypeptide (PIK3cb) shRNA in eukaryon plasmid catalyzed by PI3K in rat, then test their effects on intimal hyperplasia in transplanted vein graft. Methods One hundred and fifty SD rats were randomly divided into six groups (n=25, in each group): blank (25% Pluronic F-127), shRNA-1, shRNA-2, 1/2 (shRNA-1+shRNA-2), negative control (pGenesil-1 scramble shRNA) and positive control (wortmannin) group. The jugular vein in rats were interpositioned autologously into the common carotid artery. shRNA and 25% Pluronic F-127 were mixed and coated around the transplanted vein in three PIK3cb shRNA groups. Every 5 samples were removed according to the time point (1, 3, 7, 14 and 28 days after operation), respectively. The thickness of intima and neointima area were calculated and analyzed by computer system. The PCNA expression was detected by Western blot and SP immunohistochemistry. Results The intimal thickness of three PIK3cb shRNA groups were lower than those in the blank group and negative control group on day 3, 7, 14, 28 after operation (P<0.05); The neointima area in three PIK3cb shRNA groups (except shRNA-2 group on day 3, 7) began to decrease significantly from day one (P<0.05). The protein expression of PCNA in three PIK3cb shRNA groups on day 3 after operation were decreased compared with blank group and negative group (P<0.05). The percentage of the PCNA positive cells area in three PIK3cb shRNA groups were significantly lower than those in blank group and negative control group in each time point (Plt;0.05). There were no significant differences between blank and negative control group in different time points (Pgt;0.05). Conclusion The PIK3cb shRNA can effectively inhibit the proliferation of vascular smooth muscle cell, which may provide a new gene therapy for the prevention of vein graft restenosis after bypass grafting.
ObjectiveTo quantitate expression of microRNA-21 (miRNA-21) in gastric cancer of different tumor stages and discuss its clinical value. Method The relative expressions of miRNA-21 were quantitated in the cancer tissues, corresponding normal gastric tissues adjacent to gastric cancer, and serums of 50 gastric cancer patients received opera-tion and confirmed gastric cancer by pathology and the serums of nongastric cancer patients in Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine and its Chongming Branch from January 2015 to January 2016 by real time quantitative PCR.
ResultsThe relative expression level of miRNA-21 in the gastric cancer tissues was significantly higher than that in the normal gastric tissues adjacent to gastric cancer. Among the TNM stageⅠ, Ⅱ, Ⅲ of gastric cancer patients, the relative expression levels of miRNA-21 in the cancer tissues were 2.17 (1.48-2.90), 4.08 (2.30-4.86), 8.64 (5.82-18.20), respectively and the differences among these three stages were statistically significant (P<0.05). The relative expression level of the serum miRNA-21 in the gastric cancer patients was significantly higher than that in the nongastric cancer patients, which in the serums for stageⅠ, Ⅱ, and Ⅲpatients were 31.00 (24.60-37.15), 39.10 (28.90-39.80), 44.15 (38.95-56.68), respectively and the differences among three stages were statistically significant (P<0.05). The relative expression level of miRNA-21 in the serums and cancer tissues had a positive correlation (r=0.86, P<0.05).
ConclusionMiRNA-21 appears to have a potential association with TNM stage of gastric cancer, which cautiously suggests that it might be a potential indicator for prediction of preoperative TNM stage of gastric cancer.
Objective
The aim of this study is to review the association between long non-coding RNA (lncRNA) and papillary thyroid carcinoma (PTC).
Method
The relevant literatures about lncRNA associated with PTC were retrospectively analyzed and summarized.
Results
The expression levels of noncoding RNA associated with MAP kinase pathway and growth arrest (NAMA), PTC susceptibility candidate 3 (PTCSC3), BRAF activated non-coding RNA (BANCR), maternally expressed gene 3 (MEG3), NONHSAT037832, and GAS8-AS1 in PTC tissues were significantly lower than those in non-thyroid carcinoma tissues. The expression levels of ENST00000537266, ENST00000426615, XLOC051122, XLOC006074, HOX transcript antisense RNA (HOTAIR), antisense noncoding RNA in the INK4 locus (ANRIL), and metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in PTC tissues were upregulated in PTC tissues, comparing with the non-thyroid carcinoma tissues. These lncRNAs were possibly involved in cell proliferation, migration, and apoptosis of PTC.
Conclusion
LncRNAs may provide new insights into the molecular mechanism and gene-targeted therapy of PTC and become new molecular marker for the diagnosis of PTC.
Objective To investigate the changes of microRNA-150 ( miR-150) in peripheral blood leukocytes in sepsis patients, and their relationship with expression of immune cytokines and sepsis severity. Methods The level of mature miR-150 was quantified by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and normalized to that of control miRNA, U6, in peripheral blood leukocytes of 40 patients with sepsis, 20 patients with systemic inflammatory response syndrome ( SIRS) , and 20 normal individuals. Serum concentrations of tumor necrosis factor alpha (TNF-α) and interleukin-10 (IL-10) were measured by enzyme-linked immunoabsorbent assay in all subjects. The sequential organ failure assessment ( SOFA) score systemwas used to evaluate the severity of sepsis. The relationships between miR-150 and the white blood cell count ( WBC) , TNF-α, IL-10 and SOFA score of the sepsis patients were analyzed. Results MiR-150 was stable for at least 5 days when specimen stored at 4 ℃ and the determination of miR-150 had a broad linear detecting range ( 6. 97-6. 97 ×104 pg/ μL RNA, the lowest detecting limit: 6. 97 pg/μL RNA,r=0.999) .MiR-150 expression in the peripheral blood leukocytes in the sepsis group was significantly lower than that in the healthy control group ( Plt;0.01) , while WBC, IL-10 and IL-10/TNF-α ratio were significantly higher ( Plt;0.05) . There was no significant difference in levels of miR-150, IL-10, IL-10/TNF-α ratio, and WBC between the sepsis group and the SIRS group (Pgt;0.05) . There was no significant difference in serum concentrations of TNF-α among three groups ( Pgt;0.05) . MiR-150 expression in non-survivor sepsis patients was significantly lower than that in survivor sepsis patients (Plt;0.05) , while serum IL-10 and IL-10/TNF-αratio were significantly higher (Plt;0.01) , but there was no significant difference in serum TNF-α between the non-survivor group and the survivor group ( Pgt;0.05) . There was significantly negative correlation between miR-150 and SOFA score, TNF-α and IL-10( r=-0. 619, - 0.457, -0. 431, Plt;0.05, respectively) , but no correlation between miR-150 and WBC ( r =-0. 184, Pgt;0.05) . There was no relationship between serum TNF-α, IL-10, IL-10 /TNF-α ratio or SOFA score ( Pgt;0.05) . Conclusions MiR-150 expression in the peripheral blood specimens is significantly decreased in sepsis patients. The expression level of miR-150 not only reflect the situation of inflammatory response, but also may be used as a prognostic marker in sepsis, as it can reflect the severity of sepsis in certain degree.
Intervertebral disc degeneration is a multifactorial pathological process which is one of the leading causes of disability worldwide. The main pathological changes of intervertebral disc degeneration are the degradation of extracellular matrix, apoptosis, autophagy, senescence and inflammation. Dysregulation of microRNAs has been implicated in various pathologies, including various degenerative diseases such as disc degeneration. This article reviews the research status of microRNA in degenerative disc pathology, with emphasis on the biological mechanisms and potential therapeutic prospects of microRNA in extracellular matrix degradation, apoptosis, inflammation, and cartilage endplate degeneration.
Abstract: Objective To observe the expression changes of microRNA 1 (miRNA-1) and microRNA 21(miRNA-21) after ischemic preconditioning (IPC), ischemic postconditioning (IPO) and remote ischemic preconditioning (RIPC)in an ischemia-reperfusion rat heart model in vitro, as well as the expression of their target protein heat shock protein 70 (HSP70) and programmed cell death 4 (PDCD4), and evaluate whether miRNA are involved in endogenous cardio-protective mechanism. Methods The Langendorff-perfused Sprague-Dawley rat hearts were randomly assigned into one of the four groups, control group (CON group, n=12), ischemia preconditioning group (IPC group, n=12)?, ischemia postconditioning group (IPO group, n=12) and remote ischemia preconditioning group (RIPC group,n=12). Cardiac function was digitalized and analyzed. The expression of HSP70, PDCD4, B-cell lymphoma/leukemia-2 (Bcl-2) and Bax was detected by Western blotting. The expression of miRNA-1 and miRNA-21 was detected by real-time reverse transcriotion-polymerase chain reaction (RT-PCR). Assessment of cardiac infarct size and myocardial apoptosis was determined using triphenyltetrazolium chloride (TTC) assay and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) assay respectively. Results The expressions of miRNA-1 and miRNA-21 were up-regulated in IPC group, but the expression of miRNA-1 was down-regulated in RIPC group and IPO group (P<0.05). The expressionsof PDCD4, HSP70 and Bax were down-regulated in ‘conditioning’ groups compared with CON group (P<0.05). The expression of Bcl-2 was not statistically different among the four groups. The infarct size and the myocardial apoptosis in ‘conditioning’ hearts were significantly decreased compared with CON group (P<0.05). Conclusion The expressions of the miRNA-1 and miRNA-21 are different in IPC, RIPC and IPO groups, and their target proteins are not inversely correlated with the miRNAs in all the ‘conditioning’ groups.
Objective To summarize the relationship of diabetes and its complications with microRNA. Methods Domestic and international researches were collected by searching to summarize the role of microRNA in diabetes and its complications. Results MicroRNA could affect the secretion of insulin and interfer metabolism of gulcose in fat cells, muscle cells, and liver cells, which resulting in insulin resistance. At the same time, the microRNA also played an role in damage of vascular endothelial cells and myocardial cell in diabetes. Conclusion MicroRNA acts an important role in the process of diabetes and its complications.
ObjectiveTo summarize the domestic and abroad articles related to the research on the relation between miRNA-221/222 and thyroid cancer, and explore the important effects of miRNA-221/222 in diagnosis and treatment of thyroid cancer.
MethodsDomestic and international publications involving the relationship of miRNA-221/222 to thyroid cancer were screened and reviewed.
ResultsMiRNA-221/222 is a tumor marker with high specificity and sensitivity in thyroid cancer. It has important significance for diagnosis, treatment and prognosis of thyroid cancer.
ConclusionMiRNA-221/222 is not only related to diagnosis of thyroid cancer, but also have provided a new research direction and method for gene therapy of thyroid cancer.
ObjectiveTo summarize the research progress of long non-coding RNA (lncRNA) in the regulation of malignant biological behavior of gallbladder cancer so as to provide references for its related research.MethodThe relevant literatures about studies of lncRNA in gallbladder cancer in recent years were reviewed.ResultsThe recent studies had shown that 19 lncRNAs associated with gallbladder cancer had played the important roles in regulating tumor cell proliferation, migration, invasion, apoptosis, “sponge” miRNAs, chemoresistance, and tumor metastasis. Among them, most lncRNAs tended to have carcinogenic properties, only a few had anticarcinogenic effect. Although the research suggested the mechanism and role of lncRNA to promote or inhibit the occurrence and development of gallbladder cancer, the current research on its mechanism was still limited. In addition, some lncRNAs were found to be specifically expressed in the serum of patients with gallbladder cancer, so which were expected to become biomarkers for tumor diagnosis and prognosis.ConclusionslncRNAs associated with gallbladder cancer have carcinogenic or anticarcinogenic effect, or chemoresistance. They play potential roles in diagnosis, prognosis, and (or) treatment of tumors, but molecular mechanisms of their effects are still limited.
Objective To explore the mode and role of differential expression of circular RNAs (circRNAs) in myelodysplastic syndrome (MDS). Methods We preprocessed and analyzed the circRNA expression profile datasets GSE163386, GSE94591, and GSE81173 in the GEO (Gene Expression Omnibus) database. By using the circBank database and the ENCORI, miRDB, and miRWalk databases to predict microRNAs (miRNAs) that interacted with differentially expressed circRNAs and messenger RNAs (mRNAs), the circRNA-miRNA-mRNA axis was constructed. We retrieved miRNAs related to MDS in PubMed and further obtained competing endogenous RNA (ceRNA) networks related to MDS by taking intersections. Results Through analysis, 128 differentially expressed circRNAs were identified, 48 highly expressed, and 80 low expressed. Among differentially expressed circRNAs with multiple differences>10, 3 were upregulated and 11 were downregulated. Through analysis, 101 differentially expressed mRNA were identified, with 9 upregulated and 92 downregulated. Intersecting with the MDS related miRNAs retrieved by PubMed, we further obtained the MDS related ceRNA network, namely circRNA (has_circ_0061137)-miRNA (has-miR-16-5p)-mRNA (RUBCNL, TBC1D9, SLC16A6) and circRNA (has_circ_0061137)-miRNA (has-miR-125b-5p)-mRNA (CCR5, SLC16A6, IRF4), all of which were downregulated. Conclusion The ceRNA networks revealed in this study may help elucidate the circRNA mechanism in MDS.