Role of microRNA in autoimmune and vasculitis / 中国免疫学杂志

miRNAs are a novel class of small, non-coding RNAs that regulate the expression of multiple protein-encoding genes at the post-transcriptional level by bind to the 3'-untranslated region ( UTR) of their target messenger RNAs. Although miRNAs constitute only 3％ of the human genome, they regulate approximately 90％ of genes. In this study, we are going to introduce the role of microRNAs in autoimmune and vasculitis disease.

Clinical characteristics and GAA gene mutation in children with glycogen storage disease type II: an analysis of 3 cases / 中国当代儿科杂志

Glycogen storage disease type II (GSD II) is an autosomal recessive disorder caused by a deficiency of the lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) and can affect multiple systems including the heart and skeletal muscle. The aim of this study was to investigate three children with GSD II confirmed by GAA gene analysis and to report their clinical characteristics and gene mutations. One case was classified as infantile-onset GSD II, and two cases as late-onset GSD II. The infantile-onset patient (aged 4 months) showed no weight increase and had dyspnea, muscle hypotonia, and increased alanine aminotransferase and creatine kinase; echocardiography showed hypertrophic cardiomyopathy. The late-onset patients (aged 8 years and 13 years respectively) showed persistently elevated liver enzymes; one of them had recurrent respiratory tract infection and restrictive ventilation disorder, and the other case showed significantly increased creatase but normal electromyographic findings. Peripheral blood genetic testing for GAA gene showed six pathogenic mutations in the three cases, and the mutations c.2738C>T and c.568C>T had not been reported. Therefore, peripheral blood genetic testing for GAA gene is an effective diagnostic method.

Roles of cardiac mast cells and Toll-like receptor 4 in viral myocarditis among mice / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the role and significance of cardiac mast cells and Toll-like receptor 4 (TLR4) in the development and progression of viral myocarditis (VMC).</p><p><b>METHODS</b>Forty-eight Balb/c mice were randomly divided into a control group (n=24) and a model group (n=24). Coxsackievirus B3 was intraperitoneally injected into the model group mice to establish a VMC model. In each group, cardiac tissues were collected from 8 mice at 7, 14 and 28 days after the model was established. The cardiac tissues were stained with hematoxylin and eosin as well as Masson trichrome to observe pathological changes in cardiac tissues. The number and degranulation of cardiac mast cells at each time point were measured and evaluated by toluidine blue staining and transmission electron microscopy. The mRNA and protein expression of TLR4 in cardiac tissues was measured by RT-PCR and immunohistochemistry. In the model group, the correlation between number of cardiac mast cells and mRNA expression of TLR4 at all time points was analyzed.</p><p><b>RESULTS</b>The model group had significantly higher pathological scores of cardiac tissues than the control group at all time points (P<0.05). The myocardial collagen volume fraction in the model group at 28 days was significantly higher than in the control group at all time points and higher than in the model group at 7 and 14 days (P<0.05). At each time point, the model group had a significantly increased number of mast cells (P<0.05), and significantly increased mRNA and protein expression of TLR4 (P<0.05) compared with the control group. In the model group, the number of cardiac mast cells was positively correlated with the mRNA expression of TLR4 at all time points (R<suP>2</suP>=0.877, P<0.05).</p><p><b>CONCLUSIONS</b>Mice with VMC have significantly increased numbers of cardiac mast cells and expression of TLR4 compared with control mice at all time points, suggesting that mast cells and TLR4 may play important roles in the inflammatory response and fibrosis of VMC.</p>

Effect of HeLa cells infected with Coxsackie virus B3 on mTOR and p70S6K mRNA expression under different nutritional conditions / 中华实用儿科临床杂志

Objective To explore the effect of HeLa cells infected with Coxsackie virus B3 (CVB3) on the changes of mTOR signal pathway under different nutritional conditions.Methods The HeLa cells were cultured under conventional culture and serum starvation culture.(1) For the conventional method,the medium with 10 g/L fetal bovine serum was added for 24 h after the Hela cells were fused into 40％ to 50％,and the medium was changed on the next day,then the virus group was infected with CVB3 of 50％ tissue culture infective dose (TCID50).However,the control group was cultured by 2 g/L fetal bovine serum.(2) For the serum starvation method,HeLa cells were cultured with the medium without fetal bovine serum for 24 h.Then the virus group was infected with CVB3 of TCID50.The cells in control group were cultured by 2 g/L fetal bovine serum.Cell morphology changes were observed by inverted microscope,and the expressions of the mTOR,p70S6K mRNA were detected with Real-time PCR at 3 h,6 h,9 h,12 h,24 h respectively in both conventional culture and serum starvation groups.Results The expressions of mTOR and p70S6K mRNA were lower in the virus group than those in control group at 12 h and the 24 h (all P ＜0.05) in the conventional culture group.And the expressions of mTOR and p70S6K mRNA in the virus group were lower than those in the control group at every time points (all P ＜ 0.05) in serum starvation group.The expressions of mTOR and p70S6K mRNA in group with serum starvation virus and the control groups were higher than those in conventional culture group in all time points,but only the expressions of mTOR mRNA were significantly different between the 2 groups (all P ＜0.05),however,the expressions of p70S6K mRNA had no significant difference between the 2 groups (all P ＞ 0.05).Conclusion CVB3 may be able to down-regulate the expressions of mTOR and p70S6K mRNA.

Investigation of platelet membrane glycoprotein Ibα gene polymorphisms in children with Kawasaki disease / 中华实用儿科临床杂志

Objective To investigate the association between platelet glycoprotein Ibα (GPIbα) gene HPA-2a/b polymorphisms and the risk of Kawasaki disease (KD) and that complicated with coronary artery lesion (CAL).Methods A total of 30 patients with KD and 60 healthy controls were genotyped by polymerase chain reaction-restriction fragment length polymorphism and agarose gel electrophoresis for the HPA-2a/b polymorphism in GPIbα gene.Results For HPA-2a/b polymorphism in GPIbα gene,there was only genotype TC and CC of HPA-2a/b polymorphism in GPIbα gene in children with KD and healthy controls,and genotype TF was not found in both groups.There were no significant differences between KD patients and the controls in genotype frequencies of CC,TC and TT and allele frequencies of C and T (x2 =0.052,0.048,all P ＞ 0.05) ; also there was no significant difference between KD patients with CAL and that without CAL in genotype and allele frequencies (x2 =2.672,2.481,all P ＞ 0.05).Conclusion No association is found between HPA-2a/b polymorphism in GPIbα gene and the risk of KD or its complication of CAL in this study.

Effects of mTOR inhibitor rapamycin on Smad 3 protein and collagen type I expression in rat myocardial fibroblasts infected with coxsackie virus B 3 / 中华心血管病杂志

<p><b>OBJECTIVE</b>Mammalian target of rapamycin (mTOR) plays a central role in controlling cell proliferation, survival and growth. We investigated the role of mTOR signal transduction on viral myocarditis by observing the effect of mTOR inhibitor rapamycin on Smad 3 and collagen type I expression in rat myocardial fibroblasts infected with coxsackievirus B 3 (CVB 3).</p><p><b>METHODS</b>Primary cultured myocardial fibroblasts of SD rats infected with CVB 3 were treated with or without rapamycin. The Smad 3 and collagen type I expression of the cells were determined by RT-PCR and Western blot.</p><p><b>RESULTS</b>(1) mTOR/beta-actin ratio was dose-dependently reduced (1 nmol/L, 0.381 +/- 0.022; 10 nmol/L, 0.282 +/- 0.014; 100 nmol/L, 0.263 +/- 0.012 vs. control 1.45 +/- 0.04, all P < 0.05 vs. control) after 48 hours rapamycin treatments and time-dependently reduced after 10 nmol/L rapamycin treatment (24 h, 0.203 +/- 0.021; 48 h, 0.163 +/- 0.022; 72 h, 0.144 +/- 0.013 vs. 0 h, 0.341 +/- 0.022, all P < 0.05 vs.0 h) in CVB 3 infected myocardial fibroblasts. (2) Smad 3/beta-actin ratio of myocardial fibroblasts was significantly increased in CVB 3 infected cardial fibroblasts and this increase could be significantly attenuated by rapamycin (control, 0.63 +/- 0.06; CVB 3, 1.18 +/- 0.03; CVB 3 + Rapamycin, 0.77 +/- 0.08 by RT-PCR and 0.89 +/- 0.07, 2.27 +/- 0.13 and 0.131 +/- 0.013 by Western blot). Collagen type I/beta-actin ratio was also significantly increased by CVB 3 and this increase could be reversed by rapamycin (1.13 +/- 0.06, 1.303 +/- 0.012, 0.82 +/- 0.03 by RT-PCR).</p><p><b>CONCLUSION</b>Rapamycin can inhibit the Smad 3 and collagen type I expressions in CVB 3 infected myocardial fibroblasts suggesting that the mTOR signal pathway may play an important role in the pathogenesis of CVB 3 induced myocardial fibrosis.</p>

Effeet of rapamycin on mTOR and eIF-4E expression in coxsackievirus B3-induced rat myocardial cells / 中南大学学报(医学版)

OBJECTIVE@#To observe the effeet of rapamycin, an inhibitor of mammalian target of rapamycin (mTOR), on mTOR and eukaryotic initiation factor-4E(eIF-4E)expression in coxsac-kievirus B3 (CVB3)-induced rat myocardial cells and to investigate the role of mTOR/eIF-4E signal pathway in viral myocarditis.@*METHODS@#To construct a cell model of viral myocarditis with primary cultured myocardial cells. Myocardial cells infected by CVB3 were treated with 10 nmol/L rapamycin according to the cell toxicity test. The mTOR and eIF-4E expressions of cells were determined by RT-PCR and Western Blot.@*RESULTS@#Rapamycin inhibited the degeneration of CVB3-induced myocardial cells. Expressions of mTOR and eIF-4E mRNA or protein in CVB3-induced myocardial cells were significantly upregulated compared with the control group (P < 0.05), and rapamycin (10 nmol/L) inhibited the upregulation (P < 0.05).@*CONCLUSION@#Rapamycin can downregulate the expressions of mTOR and eIF-4E in CVB3-induced myocardial cells, suggesting that mTOR/eIF-4E signal transduction may play an important role in viral myocarditis.

Rapamycin affects eIF- 4E expression in rat myocardial fibroblasts infected by Coxsackievirus B3 / 中国当代儿科杂志

<p><b>OBJECTIVE</b>This study examined the effect of rapamycin, an inhibitor of mammalian target of rapamycin (mTOR), on eukaryotic initiation factor (eIF- 4E) expression in rat myocardial fibroblasts infected by Coxsackievirus B3 (CVB3) in order to identify the drug target for treatment of viral myocarditis.</p><p><b>METHODS</b>Primary cultured rat myocardial fibroblasts were treated with CVB3 with multiplicity of infection (MOI=0.5 PFU/cell). The experiment consisted of four groups in which the cultured rat fibroblasts cells were treated with CVB3, rapamycin (10 nM) and CVB3 + rapamycin or placebo (control). Experimental model of CVB3-infected myocardial fibroblasts was confirmed by detection of CVB3 mRNA expression with RT-PCR and observation of morphological changes of the infected cells with microscopy. eIF-4E expression was determined by both RT-PCR and Western Blot methods.</p><p><b>RESULTS</b>Morphological changes were found in the fibroblasts treated with MOI 0.5 PFU/cell of CVB3 by transmission electron microscope and the viral particles were found in the cytoplasm. CVB3 mRNA was expressed in CVB3-infected fibroblasts after 1, 2, and 3 days after infection and 2 days after passage. The gray scale values of the eIF- 4E /beta -actin in the control, the CVB3, the rapamycin and the CVB3+rapamycin groups were 0.73 +/- 0.07, 0.87 +/- 0.03, 0.32 +/- 0.03 and 0.56 +/- 0.04 respectively detected by RT-PCR, and were 0.79 +/- 0.09, 1.35 +/- 0.12, 0.55 +/- 0.04, and 0.62 +/- 0.07 respectively detected by Western blot. EIF- 4E expression in the CVB3 group was higher than that in the control group. Both the rapamycin and the CVB3+rapamycin groups had lower eIF- 4E expression than the control and the CVB3 groups.</p><p><b>CONCLUSIONS</b>CVB3 can infect myocardial fibroblasts and up-regulate the eIF- 4E expression in rat myocardial fibroblasts. Rapamycin can inhibit eIF- 4E expression and may be a potential medicine for treatment of viral myocarditis. It was suspected that mTOR/eIF- 4E signal pathway in rat myocardial fibroblasts might play an important role in the pathogenesis of viral myocarditis.</p>

Life Quality of Children with Intractable Epilepsy and Effect of Psychological Intervention on Them / 实用儿科临床杂志

Objective To investigate the life quality of children with intractable epilepsy and observe the effect of psychological intervention on them.Methods Thirty-nine children with intractable epilepsy,42 children with drug respond epilepsy and 40 healthy children were employed and tested by using of the child self-report scale of quality of life for children with epilepsy.Scores of quality of life were compared among 3 groups.Children with intractable epilepsy were divided randomly into 2 groups:psychological intervention group(19 cases)and non psychological intervention group(20 cases).Only drug treatment was given in non psychological intervention group,drug treatment and psychological intervention were given in psychological intervention group,quality of life was valuated before and 1 month after psychological intervention,then scores of quality of life were compared after and before psychological intervention in psychological intervention group,total scores of quality of life were compared between psychological intervention group after and before psychological intervention and non psychological intervention group.Results Compared with children with drug respond epilepsy and healthy children,the children with intractable epilepsy had lower scores of quality of life(Pa