Effect of overexpression of LYRM1 gene on reactive oxygen species in skeletal muscle cells / 中华实用儿科临床杂志

Objective To explore the effect of LYRM1 overexpression on production of reactive oxygen species (ROS) in skeletal muscle cells.Methods Rat myoblasts(L6) were transfected with either an empty vector or a LYRM1 expression vector.Cells were screened and the expression of LYRM1 protein in cells was identified.L6 cells were incubated in culture solution with H2-DCFDA after they were differentiated.Then fluorescence intensity of ROS in L6 was observed by fluorescence microscope,and the content of ROS was determined by flow cytometry.Results The relative fluorescence intensity of ROS in L6 overexpressing LYRM1 was 24.8933 ± 4.4574,while that in contrast cells was 13.1512 ± 0.7347,the difference between them was significant(t =24.12,P =0.00).Conclusions Overexpression LYRM1 can increase the production of ROS in skeletal muscle cells.LYRM1 overexpression may be influence the mitochondrial function and induce the mitochondrial damage of skeletal muscle cells.

Relative role of free fatty acids on expression of miR-335 in human matured adipocytes / 中华实用儿科临床杂志

Objective To explore the role of free fatty acids(FFA) on expression of miR-335 in human matured adipocytes.Methods In order to induce differentiation,confluent human pre-adipocytes (day 0) were subsequently cultured in serum-free PAM containing 50 nmol/L insulin,100 nmol/L dexamethasone,0.5 mmol/L 3-isobutyl1-methylxanthine,and 100 μmol/L rosiglitazone.Then human matured adipocytes (day 16) were treated with 1 mmol/L FFA cocktail composed of lauric acid,myristic acid,linoleic acid,oleic acid,and arachidonic acids for 4,8 and 24 hours.Meanwhile,untreated cells were collected as control group.Total RNA from these adipocytes were extracted and the levels of miR-335 expression were evaluated by real-time PCR.Results The expression of miR-335 at 4,8 and 24 hours showed no statistical significance when compared to 0 hour in untreated matured adipocytes (all P ＞ 0.05).The relative expression of miR-335 after the intervention of FFA in human matured adipocytes were 9.03 ± 0.31,9.85 ±2.41 and 11.23 ± 0.62,respectively at 4,8 and 24 hours when used with snRU6 for normalization,and there was statistical signi-ficance compared with 0 hour in control group (all P ＜ 0.05).The levels of miR-335 were 4.73 ± 0.60,5.38 ± 1.25 and 4.57 ±0.52 at the same time point when used with miR-103 for normalization,and there was statistical significance compared with 0 hour in control group (all P ＜ 0.05).Conclusions FFA exert a positive effect on the miR-335 expression in human matured adipocytes,which provide the basis for the further study about the role of miR-335 in human adipocytes.

Influence of cyanide p-trifluoromethoxyphenyl-hydrazone on insulin sensitivity and mitochondrial function in 3T3-L1 adipocytes / 中华实用儿科临床杂志

Objective To observe the effect of uncoupler of mitochondrial oxidative phosphorylation——Cyanide p-trifluoromethoxyphenyl-hydrazone (FCCP) on mitochondrial function and insulin sensitivity in mature adipocytes.Me-thods 3T3-L1 pre-adipocytes were differentiated into mature adipocytes and then induced and maintained in medium that contained the chemical uncoupler 7.5 μmol/L FCCP.Glucose uptake was determined in the adipocytes by measu-ring 2-deoxy-D-[3H] glucose uptake.Western blot was used to detect the translocation of insulin-sensitive glucose transporter 4 (GLUT4) and measure the phosphorylation and total protein contents of insulin signaling proteins such as insulin receptor substrate(IRS)-1,Akt.The mitochondrial morphology was performed by transmission electron microscope.The mitochondrial DNA (mtDNA) copy number was evaluated by real time PCR.Luciferase-based luminescence assay was used to determine cellular ATP production.The mitochondrial membrane potential(△Ψm) and reactive oxygen species(ROS) were detected by flow cytometry.Results (1) Exposure of mature adipocytes to FCCP basal glucose uptake was similar to mature adipocytes without FCCP(t =-0.07,P ＞ 0.05) ; however,the insulin-stimulated glucose uptake was significantly decreased in FCCP group (t =5.87,P ＜ 0.01).(2)FCCP decreased insulin-stimulated GLUT4 translocation to the plasmalemma and inhibition of insulin-induced phosphorylation of IRS-1 and Akt in 3T3-L1 adipocytes.(3)The size of mitochondria in FCCP-treated adipocytes was smaller than that in 3T3-L1 adipocytes without FCCP,and the morphology was condensed and abnormal.(4) mtDNA copy number in FCCP-treated adipocytes was significantly lower than that in adipocytes without FCCP(t =-1.73,P ＜ 0.001).(5) Exposure of 3T3-L1 adipocytes to FCCP significantly decreased △Ψm (t =4.83,P ＜ 0.01) and total cellular ATP production compared with cells without FCCP (t =6.08,P ＜ 0.0001),as well as the increased intracellular ROS levels (t =-6.82,P ＜ 0.01).Conclusions FCCP may impair mitochondrial morphology and mitochondrion dysfunction,and inhibited the activation of insulin-induced phosphorylation of IRS-1 and Akt in 3T3-L1 adipocytes,which suggests that FCCP-induced insulin resistance may be correlated with the FCCP induced generation of ROS in 3T3-L1 adipocytes.

Association of sleep and behavioral problems in Nanjing preschool children / 中华实用儿科临床杂志

Objective To investigate the prevalence of sleep and behavioral problems in a large sample of Nanjing preschoolers,and explore the relationship between them.Methods A total of 1327 children from 6 kindergartens of 2 districts in Nanjing,aged 3-6 years,were included in the study by using a stratified random sample survey method.Parents of these children completed a questionnaire including sleep habits and social characteristics of the children and their family.Behavioral problem scores were measured by the Achenbach children behavior checklist for children aged 1.5-5.0 years.The relationship between sleep and behavioral problems was tested using multivariate Logistic regression models to control for potentially confounding factors.Results Overall,52.68％ of the children were found to have sleep problems.The prevalence of sleep problems in boys was 56.11％,which was significantly higher than that (48.60％) in girls (P =0.006).The prevalence of total behavioral problems was 10.40％.Children with sleep problems had significantly higher prevalence and scores of total behavioral problems,internalizing syndrome and externalizing syndrome compared with those of children without sleep problems,and the differences were significant (P ＜ 0.05).In Logistic regression models,the children sleep problems were significantly contributed to total behavioral problems(OR =2.08,P ＜ 0.001).Conclusion The children sleep problems are common and as a risk factor for behavioral problems in Nanjing preschoolers.

Meta analysis of trends of simple obesity in children aged 0-7 years old in China / 中华实用儿科临床杂志

Objective To describe the change of obesity incidence over the years by evaluating simple obesity children aged 0-7 years old during 1986 to 2010 in China.Methods Relevant articles were searched for in PubMed,Cochrane Library,Chinese Science Citation Database,CBM,CNKI and Wanfang Database using the key words of "child"," overweight" and "simple obesity" in English and Chinese articles between Jan.1,1980 and Jul.31,2012.A criterion for inclusion was established based on valid criteria for diagnostic research.The eligible studies were collected and analyzed using Stata 10.0.Publication bias was tested by Begg's funnel plot.The heterogeneity test was performed at the same time.The appropriate models was used to calculate prevalence rate and 95％ CI and study gender effect among children.Results Fourteen articles were included with a total of 126 310 children (66 558 boys and 59 752 girls).The rate of child obesity was 4.30％,95％ CI:3.30％-5.40％ ;boys:4.80％,95％ CI:3.60％-6.00％ ;girls:3.70％,95％ CI:2.80％-4.60％.There was no difference between boys and girls in obesity incidence from 1986 to 1995(P ＞0.05),while differences appeared from 1996 to2010(P＜0.05).Conclusions In the 1986 to 2010 period both child obesity incidence and differences between boys and girls continued to increase.The government shall pay attention to such a situation at once and introduce polices to prevent the continuous development of the children simple obesity.

Blood lead levels during pregnancy and its influencing factors in Nanjing, China / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the blood lead levels (BLLs) in the duration of pregnancy and 6-12 weeks after delivery, and analyze the influencing factors of BLLs in healthy pregnant women.</p><p><b>METHODS</b>Pregnant women were recruited from September 2009 to February 2010 at the prenatal clinic in Nanjing Maternity and Child Health Care Hospital. Altogether 174 healthy pregnant women without pregnant or obstetric complications or abnormal pregnancy outcomes were enrolled as the gravida group, and 120 healthy non-pregnant women as the control group. BLLs during pregnancy were determined by flame atomic absorption spectroscopy.</p><p><b>RESULTS</b>BLLs in all the three pregnancy trimesters and postpartum were 59.8±24.3, 55.4±20.1, 55.9±19.7, and 67.6±17.4 μg/L, respectively, and the mean BLL in control group was 67.5±21.3 μg/L. BLLs during all the three trimesters were lower in the gravida group than in the control group (P=0.043, 0.021, and 0.028). Furthermore, occupations, nutrients supplementation, and time of house/apartment painted were associated with BLLs in pregnant women. Lead-related occupations, cosmetics use, and living in a house painted less than 1 year before are risk factors of high BLLs among pregnant women, while calcium, iron, zinc, and milk supplements are protective factors.</p><p><b>CONCLUSION</b>Supplementing calcium, iron, zinc, and milk, or avoiding contact with risk factors may help people, especially pregnant women, to reduce lead exposure.</p>

A new component of breast milk: microRNA / 中国当代儿科杂志

MicroRNA (miRNA) is a class of non-coding endogenous small molecule single strand RNA which is found in human body fluids. In recent years, miRNAs have been found in breast milk and parts of miRNAs are related to immune organ development and regulation of the immune function in infants. This article summarizes the functions of miRNA in breast milk and evidence-based clinical practice, and the differences between microRNA content and species in breast milk and cow milk. Understanding the role of miRNA can bring new opportunities for childhood nutrition research.

Detection of 9p partial trisomy using array-based comparative genomic hybridization / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect chromosomal aberrations in a child with developmental delay and speech and language disorders in order to explore the underlying genetic causes of congenital malformation, and to investigate the feasibility of array-based comparative genomic hybridization (array-CGH) for molecular genetic diagnosis.</p><p><b>METHODS</b>G-banding and array-CGH were applied to characterize the genetic abnormality in the three family members.</p><p><b>RESULTS</b>G-banding analysis revealed the affected child and the healthy mother are both carriers of inv(9)(p13q13), while the child has carried a chromosome fragment derived from chromosome 13. Array-CGH analysis indicated the derivative chromosome fragment has originated from 9p with breakpoints at around 9p13.1-p24.3.</p><p><b>CONCLUSION</b>Trisomy 9p13.1-p24.3 may be the cause of congenital malformation in the child. For its high resolution and high accuracy, array-CGH is a powerful tool for genetic analysis.</p>

Effects of maternal deficiency of folic acid during pregnancy on pulmonary development and SP-A expression in newborn rats / 中国当代儿科杂志

<p><b>OBJECTIVE</b>This study examined the effects of maternal deficiency of folic acid during pregnancy on pulmonary development and protein A (SP-A) expression in newborn rats in order to explore the possible mechanism of lung developmental disorders.</p><p><b>METHODS</b>Thirty-six adult Sprague-Dawley female rats were randomly assigned into two groups: control and study (n=18). The study and the control groups were fed with fodder containing folic acid or not respectively. Two weeks later, the female rats in the two groups copulated with normal male rats. Newborn rats were sacrificed at 1, 7 and 14 days after birth (8 pups at each time point). Lung sections were stained with hematoxylin and eosin for histological examination. SP-A expression of protein and mRNA were determined by immunohistochemistry and real-time quantitative RT-PCR, respectively.</p><p><b>RESULTS</b>The newborn rats from the study group showed damaged lung tissue structures. The mean optical density of type II cells with positive expression of SP-A decreased significantly from 1 to 14 days in newborn rats of the study group compared with the control newborn rats (P<0.05). The real-time quantitative RT-PCR showed that the expression of lung SP-A mRNA also decreased significantly from 1 to 14 days in newborn rats of the study group compared with control newborn rats (P<0.05).</p><p><b>CONCLUSIONS</b>Maternal deficiency of folic acid during pregnancy can decrease the expression of SP-A in lung tissues of newborn rats, which might lead to the disorder of lung development maturation.</p>

Resistin inhibits rat insulinoma cell RINm5F proliferation / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Resistin was thought to link the obesity to type 2 diabetes. This study aimed to investigate the effect of resistin on insulinoma cell proliferation.</p><p><b>METHODS</b>pcDNA3.1-resistin was transfected into rat insulinoma cells RINm5F. Cell proliferation was assessed by the MTT assay. The resistin and SOCS3 mRNA levels were assessed by RT-PCR. The total Akt level and the phosphorylation status were assessed by Western blot.</p><p><b>RESULTS</b>The over-expressed resistin inhibited the RINm5F cell proliferation (p<0.05). SOCS-3 expression was up-regulated by resistin over-expression (3.2 folds over the control; p<0.05). Akt phosphorylation was down-regulated by resistin over-expression (0.6 fold over the control; p<0.05).</p><p><b>CONCLUSIONS</b>Resistin impairs the rat insulinoma cell RINm5F proliferation. This might be attributed to a down-regulation of Akt level caused by increased SOCS-3 expression.</p>

Effects of infant formula containing palm oil on the nutrient absorption and defecation in infants: a meta-analysis / 中华儿科杂志

<p><b>OBJECTIVE</b>To evaluate the effects of infant formula containing palm oil on the nutrient absorption and defecation in infants.</p><p><b>METHODS</b>A search in Cochrane Library, PubMed, OVID, Springer, China National Knowledge Infrastructure, Vip Chinese Periodical Database, Wanfang Chinese Periodical Database and Chinese Bio-medicine Database was performed to identify relevant English and Chinese language articles between January 1990 and March 2009. Two reviewers independently performed data extraction and appraised using Jadad instrument. Double data were input and analyzed by software of Review Manager 4.2 recommended by Cochrane Collaboration. Intestinal nutrient absorption, electrolyte content of fecal excretion, in vivo calcium deposition, and defecation were included as the target outcomes. These outcomes were evaluated as the combined standardized mean difference (SMD) and relative risk (RR) value and 95% CI of them.</p><p><b>RESULTS</b>Thirteen articles were included. Three articles meeting inclusion criteria were analyzed for the effects between infant formula containing palmitic acid at the Sn-2 positions and palmitic acid at the Sn-1, 3 positions; five articles were analyzed for the effects between infant formula containing palmitic acid at the Sn-1, 3 and without palmitic acid; another five articles were analyzed for the effects between infant formula containing palmitic acid at the Sn-2 positions and without palmitic acid. Absorption of fat and calcium was higher, the Ca(2+) of fecal excretion was lower when the infant formula provided palmitic acid at the Sn-2 positions or without palmitic acid than that determined when formula containing palmitic acid at the Sn-1 and Sn-3 positions was given (P < 0.01). The bone mineral content (BMC) and bone mineral density (BMD) increased at 3, 6 months when the infant formula without palmitic acid as compared with using the formula containing palmitic acid at the Sn-1 and Sn-3 positions (P < 0.01). The formation of calcium soaps in stool was reduced, the BMC increased when the infant formula provided palmitic acid at the Sn-2 positions as compared with using the infant formula without palmitic acid (P < 0.01). The incidence of soft stools was higher, and the incidence of hard stools was lower when the infant formula provided palmitic acid at the Sn-2 positions or without palmitic acid than that when formula containing palmitic acid at the Sn-1 and Sn-3 positions was used (P < 0.01).</p><p><b>CONCLUSION</b>Absorption of fat and calcium was lower, the Ca(2+) of fecal excretion was higher, the BMC was reduced, the incidence of hard stools increased when the infant formula provided the palmitic acid at the Sn-1 and Sn-3 positions as compared with using formula contained palmitic acid at the Sn-2 positions or without palmitic acid. However, this conclusion should be used cautiously because of the limited quality of studies included into the analysis.</p>

Effects of NYGGF4 gene over-expression on the insulin sensitivity and secretory function of adipocytes / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effect of a new obesity-related gene NYGGF4 on the insulin sensitivity and secretory function of adipocytes.</p><p><b>METHODS</b>3T3-L1 preadipocytes transfected with either an empty expression vector (pcDNA3.1; control group) or an NYGGF4 expression vector (NYGGF4-pcDNA3.1) were cultured in vitro and differentiated into the matured adipocytes with the standard insulin plus dexamethasone plus 3-isobutyl-methylxanthine (MDI) induction cocktail. 2-deoxy-D-[3H] glucose uptake was determined by liquid scintillation counting. Western blot was performed to detect the protein content and translocation of glucose transporter 4 (GLUT4). The supernatant concentrations of TNF-alpha, IL-6, adiponectin and resistin were measured using ELISA.</p><p><b>RESULTS</b>NYGGF4 over-expression in 3T3-L1 adipocytes reduced insulin-stimulated glucose uptake. NYGGF4 over-expression impaired insulin-stimulated GLUT4 translocation without affecting the total protein content of GLUT4. The concentrations of TNF-alpha, IL-6, adiponectin and resistin in the culture medium of 3T3-L1 transfected with NYGGF4 were not significantly different from those in the control group.</p><p><b>CONCLUSIONS</b>NYGGF4 over-expression impairs the insulin sensitivity of 3T3-L1 adipocytes through decreasing GLUT4 translocation and had no effects on the secretory function of adipocytes.</p>

Regulative role of TNFalpha on STEAP4 gene in matured human adipocytes / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Human STEAP4, a novel obesity-related gene, is associated with insulin sensitivity regulation in human adipocytes. This study aimed to explore the regulative role of TNFalpha on STEAP4 gene in matured human adipocytes.</p><p><b>METHODS</b>Human preadipocytes were cultured and differentiated into matured adipocytes in vitro. Fully differentiated adipocytes (Day 17) were treated with different concentrations of TNFalpha (0, 5, 10, 25 and 50 ng/mL) for 24 hrs. Total RNA and protein were extracted from the adipocytes. Levels of STEAP4 mRNA and protein expression were determined by real-time quantitative RT-PCR and Western blot respectively.</p><p><b>RESULTS</b>Different concentrations (5, 10, 25 and 50 ng/mL) of TNFalpha treatment for 24 hrs resulted in a significant increase in the STEAP4 mRNA expression of human matured adipocytes.The maximal effect was seen in the 50 ng/mL of TNFalpha treatment group. In parallel, STEAP4 protein synthesis in matured adipocytes increased in response to TNFalpha treatment of different concentrations (5, 10, 25 and 50 ng/mL) for 24 hrs. The maximal up-regulated effect was seen in the 25 ng/mL of TNFalpha treatment group.</p><p><b>CONCLUSIONS</b>TNFalpha can up-regulate STEAP4 mRNA expression in human matured adipocytes.</p>

Construction of eukaryotic expression vector of LYRM1, a novel gene related with human obesity, and establishment of its stable transfected 3T3-L1 cell line / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector of LYRM1 and to transfect it to preadipocytes cell line 3T3-L1.</p><p><b>METHODS</b>The complete coding sequence of LYRM1 gene was amplified by RT-PCR from human omental adipose tissue and was subcloned into eukaryotic expression vector pcDNA(TM)3.1/myc-His B. The recombinant plasmid was then transfected into 3T3-L1 preadipocytes by liposome. After screening culture by G418, stable transfected 3T3-L1 cell line was established. The expression of LYRM1 protein was identified by Western blot.</p><p><b>RESULT</b>The recombinant plasmid was confirmed by PCR, restriction enzyme digestion and sequencing. The stable transfected 3T3-L1 cell line was established and the LYRM1 protein was expressed successfully.</p><p><b>CONCLUSION</b>The eukaryotic expression vector of LYRM1 has been successfully established and the stably transfected 3T3-L1 cell line also established.</p>

Resistin Binding Peptide Stimulates Basal Insulin Secretion of RINm5F Insulinoma Cells / 实用儿科临床杂志

Objective A resistin binding peptide (RBP) was selected by phage display in our previous work. Studies had shown that RBP could antagonize the role of resistin on the lipid metabolism and endocrine function of adipose tissue, but whether RBP affects the insulin secretion of pancreatic cells is still unknown. The aim of this study is to assess the effect of RBP on basal insulin secretion in RINm5F insulinoma cells. Methods The cell viability was measured by 3-[4,5-dimethyhhiazol-2-yl]-2,5-diphenyltetra-zolium bromide (MTT) cytotoxicity assay. The supernatants were assayed for insulin content by enzyme linked immunosorbent assay (ELISA). Reverse transcriptase-PCR assay and Western blotting were used to determine the expression of glucose transporter 2 (GLUT2) involved in insulin secretion. Cytosolic Ca2+, the trigger of insulin exocytosis, was analyzed with the fluorescent probe FURA-3/AM. Results RBP did no effect on the cell viability with a concentration of 10-8-10-12mol/L of 2 hours intervention. But it stimulated basal insulin secretion of RINm5F cells, accompanied by up-regulated increased expression of GLUT2 and elevated concentration of cytosolic Ca2+. Conclusion RBP could stimulate basal insulin secretion without affecting the cell viability.

Evidence of inhaled nitric oxide therapy for hypoxic respiratory failure in term and near-term infants / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the evidence-based therapy of inhaled nitric oxide (iNO) for hypoxic respiratory failure (HRF) in term and near-term infants by analyzing the literature systematically.</p><p><b>METHODS</b>The literature related to the treatment of HRF with iNO was retrieved from the following: PubMed, EMBASE, OVID, Springer and Chinese Journals Full-Text Database (CNKI). The relevant literature on randomized controlled trials (RCTs) that met the criteria was statistically analyzed by the software of Review Manager 4.2, as recommended by Cochrane Collaboration.</p><p><b>RESULTS</b>A total of 162 articles were retrieved. Fifteen met the criteria and were selected for Meta analysis (4 single center and 11 multicenter randomized trials). Meta analysis showed that 30-60 minutes iNO therapy decreased the oxygenation index (P<0.05), increased PaO2 significantly, and reduced need of extracorporeal membrane oxygenation(ECMO) (P<0.05). However, for the neonates with HRF caused by congenital diaphragmatic hernia, iNO therapy did not result in a significant reduction in the oxygenation index and death rate. There was no significant difference in the occurrence of neurodevelopmental sequelae between the iNO and control groups.</p><p><b>CONCLUSIONS</b>The currently published evidence from RCTs supports the use of iNO in term and near-term infants with HRF but except for the HRF infants caused by diaphragmatic hernia. The effect of iNO therapy on long-term neurodevelopment needs to be further studied.</p>

Effects of small interfering RNA (siRNA) against Par-4 gene on the apoptosis of human bone marrow mesenchymal stem cells / 中华儿科杂志

<p><b>OBJECTIVE</b>The prostate apoptosis response factor-4 (Par-4) gene was originally identified by differential screening for genes that are up-regulated when prostate cells are induced to undergo apoptosis. Par-4 was found to possess potent apoptotic activity in various cellular systems in response to numerous stimuli. The aim of this study was to explore the effects of small interfering RNA (siRNA) against Par-4 gene on the apoptosis of human bone marrow mesenchymal stem cells (hBMSCs) exposed to glutamate.</p><p><b>METHODS</b>Primary culture of hBMSCs was carried out and siRNAs targeted Par-4 gene (Par-4-SiRNA) were chemically synthesized. Eukaryocytic expression vector was built and were transfected into hBMSCs with liposome. After selecting with G418, the stable cell clones were treated with glutamate. The expression of Par-4 mRNA was determined by real-time PCR. The apoptosis of hBMSCs was quantified by flow cytometry. Western blotting was used to detect the protein levels of phosphorylated Akt1 (Thr308). Relative Caspase-3 activity was determined by colorimetric assay.</p><p><b>RESULTS</b>The Par-4-SiRNA-1 and Par-4-siRNA-2 could markedly down-regulate the mRNA levels of Par-4 gene in hBMSCs. With the transfections of Par-4-SiRNA-1 and Par-4-SiRNA-2, the levels of Par-4 mRNA were respectively decreased by 88% and 67%. Both Par-4-SiRNA-1 and Par-4-SiRNA-2 inhibited significantly the apoptosis of hBMSCs induced by glutamate, in which the percentages of apoptotic cells were respectively decreased to 38.80% +/- 3.97% (P < 0.01) and 45.49% +/- 4.32% (P < 0.01) from 60.30% +/- 6.82%. Western blot assays demonstrated that, glutamate down-regulated the expression of phosphorylated Akt1 proteins in hBMSCs (89.07 +/- 6.42 and 28.30 +/- 5.65, respectively, P < 0.01). However, Par-4-SiRNA-1 and Par-4-SiRNA-2 could markedly recover the down-regulation of Akt1 proteins induced by glutamate (63.56 +/- 6.75 and 45.59 +/- 4.88, respectively, P < 0.01). And the relative Caspase-3 activity which was enhanced by the treatment with glutamate (0.1428 +/- 0.0495 and 0.8616 +/- 0.1051, P < 0.01), was suppressed by Par-4-SiRNA-1 and Par-4-SiRNA-2 (0.8616 +/- 0.1051 and 0.6581 +/- 0.0555, respectively, P < 0.01).</p><p><b>CONCLUSION</b>SiRNA against Par-4 gene could inhibit the apoptosis of hBMSCs induced by glutamate, and its inhibitory effects may be mediated by the up-regulation of phosphorylated Akt1 and the suppression of the relative Caspase-3 activity.</p>

A Meta-analysis on the risk factors of perinatal congenital heart disease in Chinese people / 中华流行病学杂志

Objective To investigate the main risk factors related to the incidence of perinatal congenital heart disease (CHD) in Chinese people. Methods Results on the risk factors of CHD in 12 papers were analyzed by Meta-analysis method. The cumulative cases and controls were 3436 and 3976, respectively. The calculation methods of the combined odds ratio( OR ) and 95 % confidence interval (CI) were determined according to the homogeneity test. Results The pooled OR values of single-factor-analysis were as follows: spiritual stimulus ( 4.55 ), maternal exposures to pesticides ( 4.85 ), negative life events (5.39) during pregnancy, cold (3.18) during early pregnancy, exposure to chemical toxic substances before pregnancy. The pooled OR values of multiple-factor analysis were as follows: spiritual stimulus (4.08), exposure to chemical toxic substances ( 3.54 ), cold with fever ( 5.00 ) during pregnancy. Conclusion The main factors influencing the incidence of CHD in Chinese people were current spiritual stimulus, exposure to chemical toxic substances or pesticides, negative life events during pregnancy, cold or fever during early pregnancy, exposure to chemical toxic substances before pregnancy.

Overexpression of resistin affect 3T3-L1 adipocyte lipid metabolism / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the effect of resistin overexpression on 3T3-L1 adipocyte lipid and glucose metabolism.</p><p><b>METHODS</b>Expression vector for rat resistin gene was constructed and transfected into 3T3-L1 adipocytes. Cell differentiation and lipid accumulation was determined by Oil Red O staining. Differentiation marker genes (pref-1, C/EBPalpha, FAS) and glucose transporter 4 (GLUT4) gene mRNA expressions were evaluated by reverse transcription-PCR (RT-PCR). Triglyceride (TG) and free fatty acids (FFAs) in adipocytes were measured by colorimetric kit.</p><p><b>RESULTS</b>(1) In resistin-overexpressed adipocytes, the lipid droplets presented at the second day which was earlier than the control cells. (2) The expression of C/EBPalpha and FAS genes in resistin-overexpressed adipocytes were up-regulated and the pref-1 was down-regulated compared with that of the control cells. (3) In resistin-overexpressed adipocytes, cellular TG and FFAs levels were significantly increased (P<0.05). (4) There was no difference in the expression of GLUT4 gene between 3T3-L1 adipocytes and resistin-overexpressed adipocytes (P> 0.05).</p><p><b>CONCLUSION</b>Overexpression of resistin can affect 3T3-L1 adipocyte lipid metabolism and thereby result in obesity and insulin resistance, but have no effect on GLUT4 gene expression.</p>

A resistin binding peptide selected by phage display inhibits 3T3-L1 preadipocyte differentiation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Resistin, a newly discovered cysteine-rich hormone secreted mainly by adipose tissues, has been proposed to form a biochemical link between obesity and type 2 diabetes. However, the resistin receptor has not yet been identified. This study aimed to identify resistin binding proteins/receptor.</p><p><b>METHODS</b>Three cDNA fragments with the same 11 bp 5' sequence were found by screening a cDNA phage display library of rat multiple tissues. As the reading frames of the same 11 bp 5' sequence were interrupted by a TGA stop codon, plaque lift assay was consequently used to prove the readthrough phenomenon. The stop codon in the same 11 bp 5' sequence was replaced by tryptophan, and the binding activity of the coded peptide [AWIL, which was designated as resistin binding peptide (RBP)] with resistin was identified by the confocal microscopy technique and the affinity chromatography experiment. pDual GC-resistin and pDual GC-resistin binding peptide were co-transfected into 3T3-L1 cells to confirm the function of resistin binding peptide.</p><p><b>RESULTS</b>Three cDNA fragments with the same 11 bp 5' sequence were found. The TGA stop codon in reading frames of the same 11 bp 5' sequence was proved to be readthroughed. The binding activity of RBP with resistin was consequently identified. The expression of the resistin binding peptide in 3T3-L1 preadipocytes expressing pDual GC-resistin significantly inhibited the adipogenic differentiation.</p><p><b>CONCLUSION</b>RBP could effectively rescue the promoted differentiation of resistin overexpressed 3T3-L1 preadipocyte.</p>