Obesity is associated with the Arg389Gly ADRB1 but not with the Trp64Arg ADRB3 polymorphism in children from San Luis Potosí and León,México

This research was designed to analyze the possible associations of Arg389 Gly ADRB1 and Trp64 Arg ADRB3polymorphisms in children with obesity.A cross-sectional study included 1,046 school-age Mexican participants（6-12 years old） from the cities of San Luis Potosi and Leon.Children were classified as non-obese or obese according to their body mass index（BMI） percentile;obese children had a BMI≥95th percentile for sex and age.Biochemical data were collected.Polymorphisms were detected using TaqMan qPCR assay.A logistic regression analysis was used to calculate the risk of obesity based on genotypes.Differences were found between groups where obese children had a significant increase in systolic and diastolic blood pressure,fasting plasma glucose,insulin,HOMAIR,LDL-cholesterol,triglycerides,and lower HDL-cholesterol compared with the normal weight group（P 〈 0.05）.The distribution of allele frequency in the population was Arg = 87.4 and Gly = 12.6（Hardy Weinberg equilibrium x^2= 3.16,P = 0.07）;Trp = 81.5 and Arg= 18.5（Hardy Weinberg equilibrium x^2 = 2.2,P = 0.14） for ADRB1 and ADRB3,respectively.Even though no different frequencies of Arg389 Gly polymoiphism between groups were found（P = 0.08）,children carriers of one Gly389,ADRB1 allele had a risk for obesity of OR = 1.40（95%CI,1.03-1.90,P =0.03） after adjustment for age and gender.No other association was found for Trp64 Arg ADRB3 polymorphism.Only the Arg389 Gly ADRB1 polymorphism was associated with risk for obesity in Mexican children.

Expression and significance of fat mass and obesity associated gene and forkhead transcription factor O1 in non-alcoholic fatty liver disease

Background Non-alcoholic fatty liver disease （NAFLD） is a complex disorder and has been closely linked to obesity.The fat mass and obesity-associated （FTO） gene is a newly discovered gene related to obesity,which enhances oxidative stress and tipogenesis in NAFLD.The forkhead transcription factor O1 （FoxO1） is another important gene involved in NAFLD,which causes lipid disorders when insulin resistance appears in the liver.However,the interactions between FTO and FoxO1 during the pathogenesis of NAFLD have not been fully elucidated.This study was designed to identify the relationship between these two factors that are involved in the development of NAFLD.Methods This study includes two parts referred to as animal and cell experiments.Twelve female SPF C57BL/6 mice were fed a high-fat diet to serve as an NAFLD animal model.Aspartate aminotransferase （AST）,alanine aminotransferase （ALT）,total triglyceride （TG）,total cholesterol （TC）,alkaline phosphatase （ALP）,high-density lipoprotein （HDL）,and low-density lipoprotein （LDL） were measured.Immunohistochemical analysis was used to detect the expression and histological localization of FTO,FoxO1,and adenosine monophosphate （AMP）-activated protein kinase （AMPK）.The L02 cells were exposed to high fat for 24,48,or 72 hours.Oil red O staining was used to detect intracellular lipid droplets.Reverse transcription-polymerase chain reaction was used for analyzing the levels of FTO and FoxO1 mRNA.Results At the end of 10 weeks,ALP,ALT,AST,and LDL were significantly increased （P ＜0.01）,while TC and TG were also significantly higher （P ＜0.05）.In addition,HDL was significantly decreased （P ＜0.05）.The FTO and FoxO1 proteins were weakly expressed in the control group,but both FTO and FoxO1 were expressed significantly higher （P ＜0.01） in the experimental group,and the expression of the two factors was significantly correlated.AMPK in the high-fat group showed a low level of correlation with FTO,but not with FoxO1.Oil Red O staining results showed that the cells cultured in 50％ fetal bovine serum for 24,48,or 72 hours exhibited steatosis.FTO and FoxO1 mRNA were increased in the high-fat group compared with the normal group （P ＜0.01）.The expression levels of FTO and FoxO1 mRNA were the highest at 48 hours （P ＜0.05）.Conclusions A high-fat diet leads to higher expression of FTO,phosphorylation of FoxO1,and decreased phosphorylation of AMPK.These results suggest that the interactions between FTO and FoxO1 are closely related to the pathogenesis of NAFLD.

Association analysis of FTO gene polymorphisms and obesity risk among Egyptian children and adolescents

Obesity is a common disorder that has a significant impact on human health as it may lead to many serious diseases and sometimes morbidity.Previous genome-wide association studies(GWAS)confirmed that there is a relationship between some variants in the first intron of the fat mass and obesity associated(FTO)gene and obesity in adults and children in different ethnic groups.In our study,the association of the FTO rs9939609 and rs17817449 variants with obesity was investigated in Egyptian children and adolescents.We examined rs9939609 and rs17817449 polymorphisms in 100 control and 100 obese cases,we used the restriction fragment length polymorphism(RFLP)technique to genotype the samples.The current study showed that there were no significant differences(P>0.05)between the cases and controls in both variants of rs17817449 and rs9939609 polymorphisms.However,there were significant correlations between rs17817449 and cholesterol and between rs9939609 and LDL.In Current Study although the two variants(rs9939609 and rs17817449)didn’t show an association with obesity,but there was a correlation between the lipid profile and these two variants.

Organ-specific alterations in circadian genes by vertical sleeve gastrectomy in an obese diabetic mouse model

Current therapies for obesity and related complications have been shown to have limited benefits,including unsatisfactory weight loss and poor metabolic improvement.With recent developments in bariatric surgery,promising advancements have been made in clinical and scientific research,particularly in the management of obesity and diabetes.Vertical sleeve gastrectomy(VSG)has become increasingly popular due to its safety,simplicity,

The exploration of the correlation between the risk of obesity and the promoter methylation of PRDM16 gene

Objective To explore the association between the Cp G methylation level of positive regulatory domain containing 16(PRDM16)gene promoter and obesity or body mass index(BMI).Methods A total of 116 patients(91female adults and 25 male adults)with abdominal operation in a municipal hospital of Henan province were enrolled in this study and they were divided into

The association between common genetic variation in the FTO gene and metabolic syndrome in Han Chinese

Background Genome-wide association studies for type 2 diabetes mellitus （T2DM） identified FTO gene as a locus conferring increased risk for common obesity in many populations with European ancestry. However, the involvement of FTO gene in obesity or T2DM related metabolic traits has not been consistently established in Chinese populations. The objective of this study was to investigate the association of FTO genetic polymorphisms with metabolic syndrome （MetS） in Han Chinese. Methods We tested 41 FTO single nucleotide polymorphisms （SNPs） for association between FTO and MetS-related traits. There were a total of 236 unrelated subjects （108 cases and 128 controls）, grouped according to the International Diabetes Federation （IDF） criteria. Results Of the 41 SNPs examined, only SNP rs8047395 exhibited statistical significance （P=-0.026） under a recessive model, after Bonferroni adjustment for multiple testing （OR 1.64, 95% CI 1.11-2.42; P=-0.014）. The common distributions of this polymorphism among Chineseawith a minor allele frequency （MAF） of 36% in the control group versus 48% in the Met$ group--greatly improved our test power in a relatively small sample size for an association study. Previously identified obesity- （or T2DM-） associated FTO SNPs were less common in Hart Chinese and were not associated with MetS in this study. No significant associations were found between our FTO SNPs and any endophenotypes of MetS. Conclusions A more common risk-conferring variant of FTO for MetS was identified in Han Chinese. Our study substantiated that genetic variations in FTO locus are involved in the pathogenesis of MetS.

Molecular cloning, sequencing and expression of obese gene in the Chinese

Objective To construct the human obese(ob) cDNA clone in the Chinese, and analyze the expression of the ob gene in adipose tissue of obese, non obese subjects and nooinsulin dependent diabetes mellitus (NIDDM) Chinese patients Methods A ob cDNA clone was isolated by reverse transcription polymerase chain reaction (RT PCR) Four groups of Chinese subjects participated in the study: 1) 12 obese subjects [body mass index (BMI): 28 5±2 3? kg/m 2]; 2) 11 non obese subjects (BMI: 21 0±1 5? kg/m 2); 3) 8 obese NIDDM patients (BMI: 27 0±1 4?kg/m 2); 4) 11 non obese NIDDM patients (BMI: 21 2±1 4?kg/m 2) The expression of ob gene mRNA in abdominal subcutaneous adipose tissue was examined using RNA dot blot hybridization with a digoxigenin labeled human ob cDNA probe The hybridized signals were quantitated by densitometry Results A full human ob cDNA fragment which included a glutamine codon at +49 was obtained A base substitution (A to G) in the coding region at position 287 was found, resulting in a glutamine being replaced by an arginine Expression of the ob gene was significantly higher in Chinese obese subjects compared to non obese ones ( P <0 05), and positively correlated with the BMI No significant difference in the amount of ob mRNA was detected between non diabetic and diabetic groups at the same BMI level Conclusions We constructed a full length human ob cDNA clone. The expression of the ob gene was significantly higher in Chinese obese subjects than in non obese ones. The metabolic and hormonal changes associated with NIDDM are not the main factors regulating the expression of the ob gene.

Molecular cloning of the obese gene from Cyprinus carpio and its expression in Escherichia coli

Aiming to analyze the characteristics of the Cyprinus carpio obese gene structure and the biological activity of its expression product,we amplified the carp obese gene using reverse transcription–polymerase chain reaction from carp mesentery adipose tissue RNA.Sequence analysis revealed that it has a length of 438 nt,which encodes a 146-amino acid peptide.When nucleotide sequence and deduced amino acid sequence were compared with homologous sequences from those of humans,pigs,and rats,they displayed a fairly high degree of conservation(the homology of the nucleotide sequence was 84%,86%,and 95%,respectively,while that of the amino acid sequence was 84%,82%,and 96%,respectively,for humans,pigs,and rats).The cDNA fragment was inserted into the expression vector pET-28a,and the resulting plasmid was expressed in Escherichia coli BL21(DE3)by isopropylthiogalactoside induction.Results of sodium dodecyl sulfate–polyacrylamide gel electrophoresis analysis indicated that a fusion protein was specifically expressed in E.coli BL21(DE3).The weight of the fusion protein was about 20 kDa,and a 16-kDa protein was expressed from the carp obese gene.By gel thin-layer scanning analysis,the amount of target protein was determined to be about 20%.The purified product was found to be biologically active and to reduce the food intake and body weight of mice during tests.

Cloning of Chinese obese cDNA and its expression in E coli

Objective To obtain the sequence of Chinese obese (OB) cDNA and establish a method of leptin production in China Methods Han Chinese OB cDNA fragment was obtained by reverse transcriptase polymerase chain reaction (RT PCR) with total RNA extracted from human adipocytes and was inserted into the expressing vector pBV220 Then the constructed recombinant plasmid pBV220 OB was transformed to E coli DH5α for leptin expression The recombinant expressing system was confirmed by restriction endonuclease digestion, DNA sequencing and protein expression E coli cells were lysed by high pressure homogenization After cell membrane was extracted, the inclusion bodies were mainly renatured and purified primarily by precipitation with ammonium sulfate and gel chromatography through a Sephadex G75 column The activity of recombinant leptin was determined by its influence on the satiety and weight gain of mice Results Analysis of DNA sequence showed that Han Chinese OB cDNA included the glutamine codon at 49 The amount of recombinant leptin expressed in E coli accounted for 31%-47% of total cellular proteins From 1?L of fermentative bacteria about 40?mg of pure recombinant human leptin was isolated with a purity of being above 95% The recombinant human leptin could reduce food intake and inhibit weight gains in mice Conclusion The glutamine codon at 49 is not missing in Chinese OB gene The biologically active human leptin can be obtained by a relatively simple method of recombinant DNA technology