NLRC3 silencing accelerates the invasion of hepatocellular carcinoma cell via IL-6/JAK2/STAT3 pathway activation.

Nucleotide-binding domain, leucine-rich repeat family with a caspase activation and recruitment domain 3 (NLRC3) participates in both immunity and cancer. The aim of this study was to determine the role of NLRC3 in human hepatocellular carcinoma (HCC) and the underlying mechanisms. We collected human liver tissues from nonalcoholic steatohepatitis (NASH), HCC, and adjacent normal tissues to characterize the pattern of NLRC3 expression by real-time quantitative polymerase chain reaction and immunohistochemistry. Then, we used the HCC cell line, HuH-7, transfected with small interfering RNA to silence the NLRC3 expression. 5-Ethynyl-2'-deoxyuridine assay, scratch assay, and transwell invasion assay were used for assessing proliferation, migration, and invasion, respectively. Flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay were conducted to assess cell apoptosis. The expression of NLRC3 was reduced in human HCC tissues, compared with normal liver and nonalcoholic steatohepatitis tissues. After knocking down of NLRC3, the proliferation, migration, and invasion were increased in HuH-7 cells. And flow cytometry and TUNEL assay showed that HuH-7 cell apoptosis was suppressed after NLRC3 knockdown. As for the underlying mechanisms, knockdown of NLRC3 in HuH-7 cells was associated with the activation of Janus kinase 2/signal transducers and activators of transcription 3 (JAK2/STAT3) pathway under interleukin-6 (IL-6) stimulation. NLRC3 expression was downregulated in human HCC tissues. NLRC3 silencing in HuH-7 cells can promote the proliferation, migration, and invasion of hepatocellular carcinoma cells. JAK2/STAT3 pathway activation induced by IL-6 may be the underlying mechanism for HCC when NLRC3 expression is silenced. And the invasion of HuH-7 cells was partially suppressed by the STAT3 specific inhibitor (cryptotanshinone). Therefore, NLRC3 may play a significant role in HCC and might be a therapeutic target for the treatment of HCC.

Increased serum myeloid-related protein 8/14 level is associated with atherosclerosis in type 2 diabetic patients.

BACKGROUND: Myeloid-related protein 8/14 (MRP8/14) is a stable heterodimer formed by two different calcium-binding proteins (MRP8 and MRP14). Studies have identified that MRP8/14 regulates vascular inflammation and serves as a novel marker of acute coronary syndrome. In this study, we evaluated the correlation between serum levels of MRP8/14, hsCRP, endogenous secretory receptor for advanced glycation end-products (esRAGE) and the occurrence of coronary artery disease (CAD), or carotid intima-media thickness (IMT) when CAD was not yet developed in diabetic patients. METHODS: Serum levels of MRP8/14, esRAGE and hsCRP were measured in 375 diabetic patients. Then the results of those who had CAD were compared against who had not. Also, we investigated the associations between above-mentioned indicators and IMT of subjects without CAD in both diabetic group and non-diabetic one. RESULTS: Serum MRP8/14 was significantly higher in CAD than in non-CAD group (9.7 ± 3.6 ug/ml vs. 8.2 ± 3.0 ug/ml, P < 0.001). It was associated with severity of CAD (r = 0.16, P = 0.026). In non-CAD group, MRP8/14 was associated with IMT in patients with (r = 0.30, P < 0.001) or without diabetes (r = 0.26, P = 0.015). The areas under the curves of receiver operating characteristic for CAD were 0.63 (95% CI 0.57-0.68) for MRP8/14, 0.76 (95% CI 0.71-0.81) for hsCRP and 0.62 (95% CI 0.56 -0.67) for esRAGE. CONCLUSION: In summary, we report that diabetic patients with CAD had elevated plasma MRP8/14 levels which were also positively correlated with the severity of CAD and carotid IMT in patients without clinically overt CAD.

[Genetic determination of fast plasma glucose concentration and correlation with anthropometric indices].

OBJECTIVE: To study the genetic determination of fast plasma glucose (FPG) and correlation with its potential correlated traits, anthropometric measures and blood pressure. METHODS: Two hundred and eighteen Type 2 diabetes mellitus (T2DM) pedigrees composed of 1383 Chinese Han individuals residing in the East and South-East China were analyzed. Univariate variance decomposition analyses were used to estimate the narrow-sense heritability (h(2)) of FPG, anthropometric indices and blood pressure, and bivariate quantitative genetic analyses were used to estimate the genetic and environmental correlations between FPG and anthropometric measures or blood pressure. RESULTS: We found that FPG, blood pressure and all anthropometric indices except for waist to hip ratio were under significant genetic determination, and the h(2) was from 0.28 to 0.43. We did not find significant genetic and environmental correlation between FPG and anthropometric indices and blood pressure. CONCLUSION: The present study demonstrated that T2DM, obesity and hypertension were controlled by some genetic factors, and FPG shares little common genetic and environmental factors with obesity-related anthropometric indices and blood pressure in our Chinese sample population.

Association of vaspin gene polymorphisms with coronary artery disease in Chinese population and function study.

BACKGROUND: Visceral adipose tissue-derived serine protease inhibitor (vaspin) is a recently identified adipokine. Studies suggest it is involved in many diseases such as obesity, diabetes and coronary artery disease (CAD). This study is to investigate the association of single nucleotide polymorphisms (SNPs) in vaspin with CAD and its potential mechanisms. METHODS: A total of 1570 consecutive patients undergoing coronary angiography were enrolled and the genotypes were determined by TaqMan allelic discrimination. Serum vaspin concentrations and mRNA expression levels were determined by ELISA and RT-PCR, respectively. Reporter gene assay was performed to investigate the effect of polymorphism on vaspin promoter function. RESULTS: After multivariate analysis, allele A of rs2236242 was found as an independent determinant of CAD (OR=1.32, p=0.004). Rs35262691 in vaspin promoter was associated with serum vaspin concentration and mRNA expression in peripheral blood mononuclear cells (PBMC) though no association had been found with CAD. Reporter gene assay further confirmed that CC genotype of rs35262691 had 2.1±0.4-fold higher activities than TT genotype in facilitating gene expression. CONCLUSIONS: Our results show that the variants of vaspin gene are associated with serum vaspin levels and risk for CAD in Chinese population.

Molecular characterization and structure analysis of HPRT in a Chinese patient with Lesch-Nyhan disease.

Lesch-Nyhan disease (LND) is caused by deficiency of hypoxanthine guanine phosphoribosyltransferase (HPRT). The aim of the present study is to characterize the molecular deficiency of a clinical diagnosed Chinese patient with attenuated variant of LND. The coding region and the intron-exon boundaries of HPRT1 gene were sequenced by standard methods, and HPRT activity was assayed by HPLC method. Structure analysis was performed to estimate the consequence of the mutant of HPRT1 gene. A new mutation c.245T>G (p.Ile82Ser) was identified in this patient, and heterozygous mutation was found in the patient's mother. The activity of HPRT in the patient was completely undetectable. Structure study indicates that the mutation of p.Ile82Ser may lead to loss of hydrophobic side chain and disrupt its normal conformation of HPRT protein. It is helpful for diagnosis of LND that sequencing analysis of HPRT1 gene is performed in male infant and juvenile with hyperuricaemia and neurologic dysfunction in Chinese.

Relationship between thyroid-stimulating hormone and blood pressure in the middle-aged and elderly population.

INTRODUCTION: Hypothyroidism and subclinical hypothyroidism may be associated with hypertension and metabolic syndrome. The aim of this study was to investigate the relationship between thyroid-stimulating hormone (TSH) and blood pressure, as well as the relationship between thyroid function and insulin resistance in middle-aged and elderly Chinese. METHODS: This was a cross-sectional, community-based study. Serum TSH, fasting glucose and insulin were measured in 2,988 subjects aged 35-80 years. Logistic regression analysis was used to identify the risk factors for hypertension. Analysis of variance and multiple linear regression analysis were performed to characterise the relationship among TSH, insulin resistance and blood pressure. RESULTS: Higher serum TSH concentration was found to be an independent risk factor for hypertension in females (odds ratio 1.4, 95% confidence interval 1.02-1.93; p-value = 0.039). The female group with subclinical hypothyroidism and high normal TSH (2.5-4.8 mIU/L) were more susceptible to high blood pressure than those with low normal TSH (0.3-2.5 mIU/L) (p-value < 0.05). After adjustment for waist-hip ratio and body mass index, neither the correlation between blood pressure and homeostasis model assessment of insulin resistance (HOMA-IR) nor the correlation between TSH and HOMA-IR were found to be significant in this study. CONCLUSION: This study provides evidence that both subclinical hypothyroidism and high normal TSH are independent risk factors for hypertension in middle-aged and elderly Chinese women.

Association between serum fibroblast growth factor 21 and diabetic nephropathy.

Fibroblast growth factor 21 (FGF-21) is a new metabolic regulator with beneficial effects on lipid and glucose metabolism in animal models of diabetes mellitus. The aim of this study was to explore the relationship between FGF-21 and diabetic nephropathy in humans. Serum FGF-21 levels were determined in groups of control (n = 50) and type 2 diabetes mellitus (T2DM) patients with normoalbuminuria (n = 158), microalbuminuria (n = 68), and macroalbuminuria (n = 38) using enzyme-linked immunosorbent assay. Multiple linear regression models were used to analyze the associations between FGF-21 or other biomedical indices and urinary albumin excretion (UAE). Median serum FGF-21 levels were increased in T2DM patients compared with nondiabetic controls and were significantly higher in patients of higher UAE group. In groups of control and T2DM patients with normoalbuminuria, microalbuminuria, and macroalbuminuria, median serum (interquartile range) FGF-21 levels were 467.89 (294.59-519.56), 492.30 (354.59-640.42), 595.01 (480.49-792.31), and 665.20 (448.68-829.75) ng/L (P < .001), respectively. After adjustment for the confounders, FGF-21, fasting plasma glucose, and high-density lipoprotein cholesterol levels were found to be independently associated with UAE in diabetic patients. Serum FGF-21 level is independently correlated with UAE in T2DM patients, indicating that circulating FGF-21 may be involved in diabetic nephropathy.

The contributions of lean tissue mass and fat mass to bone geometric adaptation at the femoral neck in Chinese overweight adults.

BACKGROUND: Overweight or obese populations may have lower risk of osteoporotic fractures and higher bone mineral density (BMD), while bone strength is determined not only by bone material but also by bone structural parameters. Thus, the influence of body weight on bone geometry was examined in Chinese overweight adults. AIM: The purpose of this study was to explore how total body lean mass (TBLM) and total body fat mass (TBFM) contribute to the variation of bone geometry at the femoral neck in Chinese overweight adults. SUBJECTS AND METHODS: Bone geometric parameters including section modulus (Z), cross-sectional area (CSA), subperiosteal width (W), cortical thickness (CT) and buckling ratio (BR) were compared in 100 overweight (body mass index, BMI >/= 23) vs. 100 underweight subjects (BMI 0.055 for both sexes). CONCLUSION: Bone geometry may adapt primarily to mechanical load as represented by TBLM, but TBFM seemed to have no independent effect on bone geometry in Chinese overweight subjects.

Correlation and prediction of trunk fat mass with four anthropometric indices in Chinese males.

To increase our understanding of the relationships of trunk fat mass (FMtrunk) and four anthropometric indices in Chinese males, 1090 males aged 20-40 years were randomly recruited from the city of Changsha, China. Waist circumference (WC) and hip circumference (HC) were measured using standardized equipment, and three other anthropometric indices of BMI, waist:hip ratio (WHR) and conicity index (CoI) were calculated using weight, height, HC and WC. FMtrunk (in kg) was measured using a Hologic QDR 4500 W dual-energy X-ray absorptiometry scanner. There was an increasing trend of FMtrunk, %FMtrunk (percentage of FMtrunk) and BMI, WC, WHR, CoI in successively older age groups (e.g. the mean FMtrunk values were 4.63 (SD 2.58), 5.39 (SD 2.74), 5.93 (SD 2.82), 6.57 (SD 2.94) in four 5-year age groups, respectively). FMtrunk and %FMtrunk were significantly correlated with four anthropometric indices with the Pearson's correlation coefficients ranging from 0.25 to 0.86. Principal component analysis was performed to form three principal components that interpreted over 99.5% of the total variation of four related anthropometric indices in all age groups, with over 65% of the total variation accounted by principal component 1. Multiple regression analyses showed that three principal components explained a greater variance (R(2) 70.0-80.1%) in FMtrunk than did BMI or WC alone (R(2) 57.8-74.1%). The present results suggest that there is an increasing trend of FMtrunk and four anthropometric indices in successively older age groups; that age has important effects on the relationships of FMtrunk and studied anthropometric indices; and that the accuracy of predicting FMtrunk using four anthropometric indices is higher than using BMI or WC alone.

Genetic and environmental correlations between bone phenotypes and anthropometric indices in Chinese.

Height, weight, bone mineral density (BMD), and bone size are all influenced by genetic and environmental factors as well as interactions between them. Height and weight are often used in population studies to adjust the bone phenotypes. However, it is still unknown what proportion of genetic and environmental variability is shared between these anthropometric characteristics and the bone phenotypes. The genetic and environmental correlations between the bone phenotypes and anthropometric indices in Chinese subjects were studied by bivariate quantitative genetic analysis on a sample of 931 healthy subjects from 292 Chinese nuclear families aged from 19 to 79 years. BMD and bone size at the lumbar spine (L1-L4) and the hip of all subjects were measured by dual-energy X-ray absorptiometry. We found significant genetic correlations between weight and spine BMD, hip BMD, spine bone size and hip bone size, which were 0.50 (P<0.01), 0.45 (P<0.01), 0.36 (P=0.02), and 0.38 (P<0.01), respectively. Likewise, significant genetic correlations between height and spine BMD, spine bone size, and hip bone size were 0.30 (P=0.02), 0.54 (P<0.01), and 0.58 (P<0.01), respectively. The environmental correlations were found to be significant only between height and spine bone size (P<0.001) and weight and hip BMD (P=0.02). These results suggest the probability that the same genetic and environmental factors contribute to these different phenotypes. Moreover, when a candidate gene or genomic region is responsible for the variation of both bone phenotypes and anthropometric indices, its true genetic effect on the bone phenotypes may be lost after one has adjusted the phenotypic values with weight and height as random environmental factors. It may have implications for population studies of candidate genes that underlie the complex bone phenotypes and for the development of strategies for therapeutic application.

Genetic determination of variation and covariation of bone mineral density at the hip and spine in a Chinese population.

Bone mineral density (BMD) is a significant determinant of risk for osteoporosis. Genetic factors are known to account for a major proportion of variation of BMD in Caucasians. However, the degree of genetic determination of BMD in Chinese populations has seldom been investigated. The aim of our study was to investigate the magnitude of the genetic determination of BMD at the spine and hip, and their genetic covariation, in a population of Shanghai city in P. R. China. The subjects consisted of 44 full-sib pairs of females aged 19-43 years, 186 mother-daughter pairs, and 270 nuclear families. For BMD at the spine and hip, the values for narrow-sense heritability h2 (+/-SE) were 0.72+/-0.14 and 0.87+/-0.14, respectively, when estimated by full-sib pairs, and 0.44+/-0.07 and 0.77+/-0.07, respectively, when estimated by mother-daughter pairs. There was a significant genetic correlation r(g) (+/-SE) of BMD between the spine and hip, of 0.97+/-0.01 and 0.76+/-0.04, respectively, when estimated by full-sib pairs and mother-daughter pairs. The common household impact on BMD in our study was negligible according to the statistical estimate. We conclude that genetic factors play a major role in the determination of the variation and covariation of BMD at the spine and hip in our Chinese sample.

Estrogen receptor alpha gene relationship with peak bone mass and body mass index in Chinese nuclear families.

Estrogen receptor alpha (ER-alpha) plays an important role in mediating estrogen signaling. Studies in Caucasian populations have shown that it is involved in endocrine-related diseases such as osteoporosis and obesity. In the present study, we first used a quantitative transmission disequilibrium test (QTDT) to examine the relationship between this gene and both the osteoporosis-related phenotype bone mineral density (BMD), and the obesity-related phenotype body mass index (BMI), in 384 Chinese nuclear families. We genotyped a dinucleotide repeat marker (TA)n, and a long-range haplotype was reconstructed using this marker and two other restriction fragment length polymorphism (RFLP) markers at PvuII and XbaI loci. Although we found significant total association [allele (TA)21 with hip BMD (P=0.001), and haplotype Px(TA)21 with spine (P=0.0007) and hip (P=0.0006) BMD], the more reliable within-family associations were not significant between these phenotype pairs. No linkage signal was obtained for either spine BMD or hip BMD. We found no association or linkage between any of the three studied polymorphisms and the long-range haplotypes of the ER-alpha gene and BMI. Our study does not support an association of the ER-alpha gene with BMD and BMI in the Chinese population.

High heritability of bone size at the hip and spine in Chinese.

Bone size, an independent determinant of bone strength, is an important risk factor for osteoporotic fracture. In the present study, we investigated the magnitude of the genetic determination of bone size at the spine and hip and their genetic covariation (if any) in a population of Chinese residing in Shanghai City of P.R. China. The subjects were 50 healthy full-sib pairs of females, 188 mother-daughter pairs, and 128 husband-wife pairs selected from 401 nuclear families. Bone size (centimeters squared) was measured at the spine and hip by dual-energy X-ray absorptiometry (DEXA). The narrow-sense heritabilities h2 (SE) of bone size at the spine and hip were 0.63 (0.14) and 0.45 (0.14) respectively when estimated by full-sib pairs, and 0.60 (0.07) and 0.69 (0.07) respectively when estimated by mother-daughter pairs. Marginally significant genetic correlation was observed between the spine and hip bone size. The significantly and moderately high h2 values for bone size demonstrated in this study warrant a subsequent genetic study to search for the genes or genomic regions underlying the phenotype in Chinese.

Parathyroid hormone gene with bone phenotypes in Chinese.

Osteoporosis is a common disorder afflicting old people. The parathyroid hormone (PTH) gene is involved in bone remodeling and calcium homeostasis, and has been considered as an important candidate gene for osteoporosis. In this study, we simultaneously tested linkage and/or association of PTH gene with bone mineral density (BMD) and bone mineral content (BMC), two important risk factors for osteoporosis. A sample of 1263 subjects from 402 Chinese nuclear families was used. The families are composed of both parents and at least one healthy daughter aged from 20 to 45 years. All the subjects were genotyped at the polymorphic BstBI site inside the intron 2 of the PTH gene (a nucleotide substitution of G to A at the position +3244). BMD and BMC were measured at the lumbar spine and the hip region via dual-energy X-ray absorptiometry (DXA). Using QTDT (quantitative trait transmission disequilibrium test), we did not find significant results for association or linkage between the PTH gene and BMD or BMC variation at the spine or hip. Our data do not support the PTH gene as a quantitative trait locus (QTL) underlying the bone phenotypic variation in the Chinese population.