MicroRNA-23a knockdown attenuates angiotensin Ⅱ induced hypertrophy in rat H9c2 cells via activating PTEN and AMPK pathway / 中华心血管病杂志

Objective: To investigate if microRNA (miR) -23a knockdown could attenuate angiotensin Ⅱ(AngⅡ) induced cardiac hypertrophy by activating phosphatase and tensin homolog deleted on chromosome ten(PTEN) and AMP-activated protein kinase(AMPK) pathway. Methods: Rat H9c2 cells were cultured in DMEM high glucose medium and put in 5% CO(2) incubator at 37 ℃(normal group). After 48 hours of culture, H9c2 cells were stimulated with 10 nmol/L AngⅡ to establish cell hypertrophy model (AngⅡgroup). The H9c2 cells were inoculated in a 6-well cell culture plate and cultured in an incubator at 37 ℃. When the confluence degree of cell growth was about 70%, the cells were transfected with different reagents, and 24 hours after transfection, 10 nmol/L AngⅡ was used to interfere with the cells. The H9c2 cells were divided into different groups according to the reagents, namely AngⅡ+anti-miR group(transfected with miR-23a inhibitor), Ang Ⅱ+NC group(transfected with miR-23a inhibitor negative control), Ang Ⅱ+anti-miR+si-PTEN group(cotransfected with miR-23a inhibitor and PTEN small interference RNA(siRNA)), and AngⅡ+anti-miR+si-NC group(cotransfected with miR-23a inhibitor and PTEN siRNA negative control). The surface area of single cell was measured by Image J software.The mRNA expression levels of α-actin 1 (ACTA1) and β-myosin heavy chain (β-MHC) and miR-23a were detected by quantitative real-time PCR(qRT-PCR). The expression levels of PTEN and AMPK signal pathway related proteins were detected by Western blot. In order to verify whether miR-23a targets PTEN gene, double luciferase reporter gene experiment was performed. The luciferase reporter gene vector recombinant plasmids of wild type pGL-WT-PTEN and mutant pGL-MUT-PTEN were constructed and prepared after normal sequencing. H9c2 cells was inoculated into 24-well cell culture plate and cultured overnight in 37 ℃ incubator. The cells were co-transfected with miR-23a mimic or miR-23a mimic negative control and wild type or mutant reporter gene recombinant plasmid. Forty-eight hours after transfection, firefly luciferase activity and sea kidney luciferase activity were measured, and the ratio of them was recorded as relative luciferase activity. Results: Compared with the normal group, the cell surface area, the mRNA expression levels of ACTA1, β-MHC and miR-23a were significantly higher, while the protein expression levels of PTEN and p-AMPK were significantly lower in the Ang Ⅱ group(all P<0.05). The results of double luciferase reporter gene assay showed that the relative luciferase activity of cells co-transfected with miR-23a mimic and wild-type reporter gene recombinant plasmid was lower than that of miR-23a mimic negative control (P<0.05), and PTEN served as the target gene of miR-23a. In AngⅡ+anti-miR group the mRNA expression levels of miR-23a, ACTA1 and β-MHC were lower, and the cell surface area was smaller, while the protein expression levels of PTEN and p-AMPK were higher than that in AngⅡ group and AngⅡ+NC group(all P<0.05). Compared with AngⅡ+anti-miR group, the cell surface area was bigger, the expression of ACTA1 and β-MHC mRNA was up-regulated, and the protein expression levels of PTEN and p-AMPK were down-regulated in Ang Ⅱ+anti-miR+si-PTEN group(all P<0.05). Conclusion: Inhibition of miR-23a can attenuate Ang Ⅱ-induced hypertrophy in H9c2 cells through targeting PTEN and activating AMPK signaling pathway.

BioinformaticsAnalysisofProstaticCarcinomaViaBig-Data / 中山大学学报(医学科学版)

@#【Objective】 The two databases，GEO（gene expression omnibus，GEO）and TCGA（the cancer genome alas ，TCGA），were analyzed using bioinformatics methods to screen differentially expressed genes associated and their related regulatory networks in prostate carcinoma. 【Methods】 The prostate carcinoma gene expression chip data （GSE46602 ，GSE55945） downloaded from the GEO database were integrated into the RNA- seq data of the TCGA database. And the differentially expressed genes analysis was performed using GEO2R and the edgeR package of R software to extract common significant differentially expressed genes. The clusterProfiler package of R software was used to enrich the GO（gene ontology ，GO）function enrichment analysis and KEGG（kyoto encyclopedia of genes and genomes， KEGG）pathway analysis. Differentially expressed genes were further constructed into a protein-protein interaction（PPI） network to screen out key genes for regulatory protein expression in prostate carcinoma. Gene analysis results were combined with TCGA clinical follow-up data to analyze the clinical prognostic value of key node genes. 【Results】A total of 278 significant differentially expressed genes were extracted，of which 178 genes were down- regulated and 100 genes were up-regulated. These genes were closely associated with the function and pathway enrichment such as the regulation of proliferation of epithelial cells，metabolism of benzene- containing compounds，the glutathione metabolism，and focal adhesion. The protein-protein interaction network analysis revealed three key protein expression modules and 12 key node genes. Among these key genes，EDN3（endothelin-3），EDNRB（endothelin receptor B）and AMACR（alpha-methylacyl- coa racemase）were closely related to the survival rate of prostate cancer patients. 【Conclusion】Through bioinformatics analysis of gene chip and RNA-seq data in prostate carcinoma，we found that EDN3，EDNRB and AMACR may play an important role in the occurrence and development of prostate carcinoma.

Correlation between COMT gene polymorphism and early cerebral vasospasm after aneurismal subarachnoid hemorrhage / 第二军医大学学报

Objective To analyze the correlation between catechol-O-methyltransferase (COMT) gene polymorphism and cerebral vasospasm (CVS) in early period after subarachnoid hemorrhage (SAH). Methods The clinical data of 167 patients with spontaneous SAH, who were treated in our hospital from Jan. 2008 to Dec. 2008, were collected for this study. COMT genotyping was performed by means of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The degree of CVS was identified by transcranial Doppler (TCD). Hunt-Hess classification was used to evaluate the severity of the patients’ condition. The bleeding amount was evaluated by means of Fisher classification of head CT.-£2 test (SPSS13.0 software) and logistic regression were adopted to analyze the correlation of COMT gene polymorphism and other clinical data with early CVS after SAH. Results The distribution of each allele matched with Hardy-Weinberg law and the research samples were heredity equilibrium population. Early CVS incidence of patients with COMT A allele was significantly higher than those with COMT G allele (51.7% vs 38.5%, P < 0.01). Early CVS incidence of patients with COMT A/A genotype was significantly higher than those with COMT G/G genotype (66.7% vs 35.9%, P < 0.05). Univariate logistic regression demonstrated that COMT A allele, A/A genotype and Grade 3-5 of Hunt-Hess classification were all associated with early CVS. After adjustment of general information, further multivariate logistic regression demonstrated that COMT A allele, A/A genotype were the risk factors of early CVS after SAH. Conclusion COMT A allele and A/A genotype might be risk factors of early CVS after SAH. Conclusion COMT A allele and A/A genotype might be risk factors of early CVS after SAH.

Correlation analysis of increased blood glucose and insulin resistance after traumatic brain injury in rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the pattern of the alterations of blood glucose, insulin and insulin sensitivity after traumatic brain injury in rats, and verify the occurrence of insulin resistance after the injury.</p><p><b>METHODS</b>Based on Feeney's model of brain injury, the blood glucose and insulin concentration of the dogs measured 30 min before and at 6, 12, 24, 48, 72 and 120 h after injury. BG60-120, GIR60-120, and insulin sensitivity index (ISI) reflecting the insulin sensitivity were measured at 6, 24, 48, and 72 hours following severe traumatic brain injury using euglycemic-hyperinsulinemic clamp.</p><p><b>RESULTS</b>Both the blood glucose and insulin concentration increased markedly in rats following moderate and severe brain injury. BG60-120 increased markedly, and GIR60-120 and ISI decreased significantly 6, 24, 48, and 72 h after severe brain trauma as compared with those of the sham operation group. Blood glucose concentration of rats following severe injury was positively correlated with insulin concentration and BG60-120 at the corresponding time points, but negatively with GIR60-120 and ISI.</p><p><b>CONCLUSION</b>Both the blood glucose and insulin concentration increase markedly in rats following severe brain injury. Increased blood glucose even in the presence of high-level insulin is due to acute insulin resistance occurring after traumatic brain injury.</p>

Advances in the investigation of biological effect and surface modification of dendrimers as drug (gene) delivery systems / 中国医学科学院学报

Dendrimers are highly branched macromolecules that have attractive nano-sized architectures. It seems that they can enter various cells easily because of their unique nanostructures and chemical properties, which make them to be one of important candidates of non-virus carriers for drug delivery or gene therapy. However, the understanding of cytotoxicity and related mechanisms of dendrimers are still limited. In recent years there has been rapid increases of researches regarding the biological effects of dendrimers, including the interactions of dendrimers to cells, transport mechanisms, intracellular distribution and biodistribution in vivo, as well as improvement of biocompatibility of dendrimers by surface engineering. In this paper, recent advances in the investigations of biological effect and surface modification for the dendrimers as drug or gene delivery systems were reviewed.

Analysis of the cause of hemorrhage after MPCNL and its interventional treatment / 中华泌尿外科杂志

Objective To analyze the cause of delayed hemorrhage after minimally invasive percu- taneous nephrolithotomy(MPCNL),and to summarize the experience in the interventional treatment of severe bleeding after MPCNL by superselective arteriolar embolization.Methods The clinical data of 3812 cases of MPCNL from June 1998 to July 2004 were reviewed.Of them,12 patients(11 men and 1 woman;mean age,45 years)who developed severe hemorrhage after MPCNL were identified.The cause of hemorrhage and the treatment results were analyzed.Results The rate of delayed hemorrhage after MPCNL was 0.31% (12/3812).The mean time to onset of severe bleeding was 10 d after MPCNL.Renal arteriography was per- formed in all 12 patients,showing 5 arteriovenous fistulas and 7 false aneurysms.Superselective arteriolar em- bolization for hemostasis was performed in all 12 cases.All these vascular abnormalities were successfully treated by superselective embolization.Follow-up showed that the hematuria disappeared and renal function recovered well.Conclusions Severe hemorrhage following MPCNL is a rare complication,the incidence of which is significantly lower than that of conventional PCNL.The cause is mainly the arteriolar injury of re- nal puncture passage.Superselective embolization provides effective control of bleeding and currently consti- tutes the treatment of choice based on our experience.