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Cytokines play a critical role in the pathogenesis and development of cardiovascular diseases. However, data linking cytokines to risk and severity of acute coronary syndrome (ACS) are still limited. We measured plasma profile of 280 cytokines using a quantitative protein microarray in 12 ACS patients and 16 healthy controls, and identified 15 differentially expressed cytokines for ACS. Osteopontin, chemokine ligand 23, brain derived neurotrophic factor and C-reactive protein (CRP) were further validated using immunoassay in two independent case-control studies with a total of 210 ACS patients and 210 controls. We further examined their relations with incident ACS among 318 case-control pairs nested within the Dongfeng-Tongji cohort, and found plasma osteopontin and CRP concentrations were associated with incident ACS, and the multivariable-adjusted odds ratio (95% confidence interval) was 1.29 (1.06–1.57) per 1-SD increase for osteopontin and 1.30 (1.02–1.66) for CRP, respectively. Higher levels of circulating osteopontin were also correlated with higher severity of ACS, and earlier ACS onset time. Adding osteopontin alone or in combination with CRP modestly improved the predictive ability of ACS beyond the Framingham risk scores. Our findings suggested that osteopontin might be a biomarker for incident ACS, using osteopontin adds moderately to traditional cardiovascular risk factors.

Objective: To study the expression quantity of serum miR-146a and miR-146b in patients with acute cerebral infarction before and after the intervention of rosuvastatin and its correlation with toll-like receptor 2 (TLR2) and TLR4 signaling pathways. Methods: A total of 65 patients with acute cerebral infarction treated in our hospital from December 2015 to August 2016 were selected for prospective study. They were treated with lipid-lowering rosuvastatin, and peripheral blood samples were collected at 8th week before and after treatment, respectively. Serum was separated and expression quantity of miR-146a and miR-146b and contents of TNF-α, interleukin (IL)-1β, IL-6 and IL-17 were determined. Peripheral blood mononuclear cells were isolated and fluorescence intensities of TLR2, TLR4, myeloid differentiation primary response gene 88 (MyD88), interleukin-1 receptor-associated kinase 1 (IRAK-1) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) were measured. >Results: At 8th week of intervention of rosuvastatin, expression quantity of serum miR-146a [(0.762 ± 0.092) vs. (0.346 ± 0.053)] and miR-146b [(0.714 ± 0.088) vs. (0.317 ± 0.047)] in patients with acute cerebral infarction was significantly higher than those before the intervention. Fluorescence intensities of peripheral blood mononuclear cells such as TLR2 [(10.34 ± 1.27) vs. (16.94 ± 1.94)], TLR4 [(11.37 ± 1.54) vs. (24.35 ± 3.26)], IRAK [(9.34 ± 0.92) vs. (15.32 ± 1.82)], MyD88 [(4.42 ± 0.56) vs. (9.41 ± 1.03)] and NF-kB [(6.65 ± 0.78) vs. (13.49 ± 1.76)] and contents of inflammatory factors such as TNF-α [(64.26 ± 8.29) μg/L vs. (106.39 ± 13.84) μg/L], IL-1β [(37.91 ± 5.24) μg/L vs. (64.23 ± 8.33) μg/L], IL-6 [(34.28 ± 4.85) ng/L vs. (82.46 ± 11.97) ng/L] and IL-17 [(56.75 ± 7.49) ng/L vs. (98.31 ± 11.36) ng/ L] of serum were all significantly lower than those before the intervention. Expression quantity of serum miR-146a and miR-146b had a negative correlation with fluorescence intensities of TLR2, TLR4, IRAK, MyD88 and NF-kB. Fluorescence intensities of TLR2 and TLR4 in peripheral blood mononuclear cells had a positive correlation with contents of TNF-α, IL-1β, IL-6 and IL-17 in serum. >Conclusions: Treatment with rosuvastatin can up-regulate the expression quantity of serum miR-146a and miR-146b in patients with acute cerebral infarction and further inhibit the secretion of IRAK, MyD88, NF-kB, TNF-α, IL-1β, IL-6 and IL-17 mediated by TLR2 and TLR4.

Objective To analyze the serum levels of miR-126, miR-146a and its relationship with infarction area, severity of disease and inflammatory reaction degree in patients with acute cerebral infarction (ACI). Methods A total of 75 cases with ACI treated in our hospital from April 2014 to October 2015 and 80 healthy cases were respectively selected as ACI group and control group for retrospective study. Patients' clinical data were collected, and the serum levels of miR-126, miR-146a, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-6 were detected. Results Serum contents of miR-126 (0.286 ± 0.078 vs . 1.000 ± 0.169) and miR-146a (0.337 ± 0.084 vs . 1.000 ± 0.158) in patients of ACI group were significantly lower than those of control group. Contents of IL-1β [(68.4 ± 10.3) vs . (22.7 ± 5.8) ng/L], TNF-α [(126.9 ± 22.4) vs . (49.6 ± 8.4) ng/L] and IL-6 [(89.3 ± 14.7) vs . (34.8 ± 5.9) ng/L] were obviously higher than those of control group. The bigger the infarction area was, the more severer the degree of nerve defect could be. The lower the serum levels of miR-126, miR-146a were, the higher the levels of TNF-α, IL-1β, IL-6 could be. Levels of miR-126 and miR-146a were negative correlation with levels of TNF-α, IL-1β and IL-6. Conclusions An abnormal decrease in serum levels of miR-126 and miR-146a in patients with ACI was closely related to the severity of disease. Through regulating the generation of inflammatory factors TNF-α, IL-1β and IL-6, miR-126 and miR-146a may get involved in the changes of cerebral infarction condition.

Background: Smoking is a risk factor for many human diseases. DNA methylation has been related to smoking, but genome-wide methylation data for smoking in Chinese populations is limited. Objectives: We aimed to investigate epigenome-wide methylation in relation to smoking in a Chinese population. Methods: We measured the methylation levels at > 485,000 CpG sites (CpGs) in DNA from leukocytes using a methylation array and conducted a genome-wide meta-analysis of DNA methylation and smoking in a total of 596 Chinese participants. We further evaluated the associations of smoking-related CpGs with internal polycyclic aromatic hydrocarbon (PAH) biomarkers and their correlations with the expression of corresponding genes. Results: We identified 318 CpGs whose methylation levels were associated with smoking at a genome-wide significance level (false discovery rate < 0.05), among which 161 CpGs annotated to 123 genes were not associated with smoking in recent studies of Europeans and African Americans. Of these smoking-related CpGs, methylation levels at 80 CpGs showed significant correlations with the expression of corresponding genes (including RUNX3, IL6R, PTAFR, ANKRD11, CEP135 and CDH23), and methylation at 15 CpGs was significantly associated with urinary 2-hydroxynaphthalene, the most representative internal monohydroxy-PAH biomarker for smoking. Conclusion: We identified DNA methylation markers associated with smoking in a Chinese population, including some markers that were also correlated with gene expression. Exposure to naphthalene, a byproduct of tobacco smoke, may contribute to smoking-related methylation. Citation: Zhu X, Li J, Deng S, Yu K, Liu X, Deng Q, Sun H, Zhang X, He M, Guo H, Chen W, Yuan J, Zhang B, Kuang D, He X, Bai Y, Han X, Liu B, Li X, Yang L, Jiang H, Zhang Y, Hu J, Cheng L, Luo X, Mei W, Zhou Z, Sun S, Zhang L, Liu C, Guo Y, Zhang Z, Hu FB, Liang L, Wu T. 2016. Genome-wide analysis of DNA methylation and cigarette smoking in Chinese. Environ Health Perspect 124:966–973; http://dx.doi.org/10.1289/ehp.1509834

Spinocerebellar ataxia (SCA) is a group of genetic diseases of the nervous system with genetic and clinical heterogeneity. SCA is often caused by an expanded CAG repeat sequence in the encoding protein. Genetic testing is necessary to diagnose and classify the types of SCA. Next‑generation DNA sequencing usually generates a high error rate for insertion or deletion mutations, so it is unhelpful for classifying the types of SCA. In the present study, a Chinese SCA pedigree was preliminarily diagnosed with SCA1 using polymerase chain reaction (PCR) amplification. The propositus and his three younger siblings were diagnosed with SCA1 as a result of the identification of the length of the expanded CAG repeat sequence in the ATXN1 gene performed using Sanger sequencing. The current study presents a convenient and efficient method to identify causative mutations for polyglutamine SCA using PCR amplification followed by Sanger sequencing.

Rationale: Acute coronary syndrome (ACS) is a leading cause of death worldwide. Immune functions play a vital role in ACS development; however, whether epigenetic modulation contributes to the regulation of blood immune cells in this disease has not been investigated. Objective: We conducted an epigenome-wide analysis with circulating immune cells to identify differentially methylated genes in ACS. Methods and Results: We examined genome-wide methylation of whole blood in 102 ACS patients and 101 controls using HumanMethylation450 array, and externally replicated significant discoveries in 100 patients and 102 controls. For the replicated loci, we further analyzed their association with ACS in 6 purified leukocyte subsets, their correlation with the expressions of annotated genes, and their association with cardiovascular traits/risk factors. We found novel and reproducible association of ACS with blood methylation at 47 cytosine-phosphoguanine sites (discovery: false discovery rate P + T cells, CD4 + T cells, and B cells. Blood methylation of 26 replicated cytosine-phosphoguanine sites showed significant correlation with expressions of annotated genes (including IL6R , FASLG , and CCL18 ; P −4 ), and differential gene expression in case versus controls corroborated the observed differential methylation. The replicated loci suggested a role in ACS-relevant functions including chemotaxis, coronary thrombosis, and T-cell–mediated cytotoxicity. Functional analysis using the top ACS-associated methylation loci in purified T and B cells revealed vital pathways related to atherogenic signaling and adaptive immune response. Furthermore, we observed a significant enrichment of the replicated cytosine-phosphoguanine sites associated with smoking and low-density lipoprotein cholesterol ( P enrichment ≤1×10 −5 ). Conclusions: Our study identified novel blood methylation alterations associated with ACS and provided potential clinical biomarkers and therapeutic targets. Our results may suggest that immune signaling and cellular functions might be regulated at an epigenetic level in ACS.

Coronary heart disease (CHD) is a disease resulting from the interaction between genetic variations and environmental factors. Zinc finger homeobox 3 (ZFHX3) is a transcription factor and contains a poly-glutamine tract in a compositionally biased region that is encoded by exon 9, containing a cluster of CAG and CAA triplets followed by the polymorphic CAA repeats: (CAG)2(CAA)2(CAG)3CAACAG(CAA)nGCA. Thus, nine successive glutamine residues precede the poly-glutamine tract, encoded by the polymorphic CAA repeats. The aim of this study was to investigate the association of the CAA repeat polymorphism in exon 9 of the ZFHX3 gene with the risk of CHD in a Chinese population. The CAA repeat polymorphism was determined by polymerase chain reaction followed by DNA sequencing in 321 CHD patients. Genotype frequencies were compared using the non-parametric mood median test. Four alleles of CAG(CAA)10GCA, CAG(CAA)8GCA, CAG(CAA)9GCA, and CAG(CAA)11GCA were found in Chinese CHD patients in exon 9 of the ZFHX3 gene. The CAG(CAA)10GCA was a major allele (95.95%), and the CAG(CAA)8GCA was a minor allele (3.58%). The CAG(CAA)9GCA and CAG(CAA)11GCA were rare alleles (0.31% and 0.16%). The CAG(CAA)10GCA allele encodes a poly-glutamine tract of 19 residues. Importantly, the CHD patients homozygous for the CAG(CAA)10GCA allele had a higher risk of CHD, compared to the heterozygous patients carrying a CAG(CAA)8GCA allele. Moreover, the CAG(CAA)10GCA allele was significantly associated with hypertension, diabetes mellitus, or dyslipidemia (P0.05). Thus, the CAA repeat polymorphism in exon 9 of the ZFHX3 gene contributes to the CHD susceptibility in the Chinese population.

Coronary heart disease (CHD) is a leading cause of morbidity and mortality around the world and has both genetic and environmental precipitants. Genetic factors are significant in determining the level of risk factors in individuals. Variants in ZFHX3 gene are associated with atrial fibrillation in individuals of European ancestry. The aim of this study was to analyze the polymorphisms in the compositionally biased region of the ZFHX3 gene in patients with coronary heart disease in a Chinese population, and to explore their associations with coronary heart disease. We recruited 278 CHD patients and 358 age and sex matched healthy controls in a Chinese Han population, polymorphisms in the compositionally biased region of the ZFHX3 gene were determined by polymerase chain reaction followed by DNA sequencing. The genotype frequencies were calculated, and statistical analysis was performed using the non-parametric mood median test. A complex insertion/deletion polymorphism was identified in the compositionally biased region of the ZFHX3 gene in a Chinese population. Six common genotypes (GGC)4GGTGGCAGT(GGC)4GGT(GGC)8, (GGC)4GGTGGCAGT(GGC)5GGT(GGC)8, (GGC)4GGTGGCAGT(GGC)5GGT(GGC)7, (GGC)2GGTGGCAGT(GGC)5GGT(GGC)10, (GGC)4GGTGGCAGT(GGC)5GGT(GGC)5, and (GGC)4GGT(GGC)8 were found in both CHD patients and healthy controls, there was no significant difference in the six genotype frequencies between CHD patients and healthy controls. Rare genotypes (GGC)4GGTGGCAGT(GGC)2GGT(GGC)2GGT(GGC)6, (GGC)4GGTGGCAGT (GGC)8, (GGC)4GGTGGCAGT(GGC)(3)GGT(GGC)8, and (GGC)6GGT(GGC)8 were only identified in healthy controls. Rare genotypes (GGC)4GGTGGCAGT(GGC)4GGT(GGC)5, (GGC)4GGTGGCAGT(GGC)4GGT(GGC)4, and (GGC)4GGTGGCGGT(GGC)6 were only found in CHD patients. The compositionally biased region of the ZFHX3 gene contains a poly-Gly sequence. A complex insertion/deletion polymorphism exists in this region in a Chinese population, clinical significance of some rare genotypes should be explored for CHD in the future.

To analyze the chromosome aberration in a full-term male neonate with low birth weight, and to explore the possible causes for growth retardation in intrauterine development for the neonate.Genomic DNA was extracted from peripheral leukocytes of the neonate. Detection of genomic DNA copy number gain and loss was performed using microarray comparative genomic hybridization. Chromosome karyotype was obtained from cultured lymphocytes for the neonate and his parents in order to identify the origin of chromosome aberration.Gain of 10q25.2--qter (22 Mb) was observed in the full-term neonate with low birth weight. In addition, one chromosomal region, 15q26.2--qter (5 Mb) was lost. The karyotype of the neonate was 46, XY, -15, +der(15), t(10;15)(q25;q26)pat.The full-term neonate with low birth weight had a partial trisomy of 10q25.2--qter with a partial monosomy of 15q26.2--qter, both of them may contribute to the growth retardation in intrauterine development for the neonate case.

Background Ageing is related to the risk of many diseases and is affected by genetic, epigenetic, and environmental factors. Exposure to the ubiquitous pollutant polycyclic aromatic hydrocarbons (PAHs) is known to cause health damage, but the relation between such exposure with methylation ageing has not been studied. We undertook an association analysis of exposure to PAHs and DNA methylation ageing, a novel and promising ageing marker. Methods We built a model of methylation age and defined two ageing indicators (ageing rate, calculated as ratio of methylation to chronological age; and Δage, calculated as methylation minus chronological age) on the basis of genome-wide methylation data of blood in five panels of Chinese individuals (N=596). Participants included 137 coke oven workers (aged 26–60 years) who were chronically and occupationally exposed to high levels of PAHs; 101 patients with acute coronary syndrome (aged 40–86 years) from Wuhan, China; 97 patients with this syndrome (aged 38–83 years) from Guangdong, China; 162 healthy individuals (aged 26–79 years) from Wuhan; and 99 healthy individuals (aged 37–80 years) from Zhuhai, China. Exposure to individual PAHs was assessed by ten urinary PAH metabolites. We examined the relations between PAH exposure and the two ageing indicators, and investigated the potential CpGs mediating the association. Written informed consent was obtained from all participants. Our study was approved by the ethics committee of Tongji Medical College, Wuhan, China. No study subjects were from clinical trials. Findings Coke oven workers, who were exposed to high levels of PAHs at work (mean nature-log-transformed total urinary PAHs −1·909 [SD 0·461]), had significant age acceleration (2·7% increase in ageing rate [p=0·002] and 1·218-year increase in Δage [p=0·01] compared with all other panels). Urinary 3-OH-phenanthrene and 9-OH-phenanthrene were independently and significantly associated with the two ageing indicators in our non-disease panels. One unit increase of nature-log-transformed 9-OH-phenanthrene was associated with a 1·8% increase (p=0·0029) in ageing rate and a 1·1-year increase (p=0·0002) in Δage, whereas for 3-OH-phenanthrene the increase was 1·3% (p=0·0050) for ageing rate and 0·57 year (p=0·0109) for Δage. Methylation at several ageing-related and PAH-related CpGs (annotated on ELOVL2 , KLHL35 , FHL2 , BOK , PDP1 , and OTUD7A ) was reported to mediate the effect of PAH exposure on methylation ageing. Interpretation Our study revealed a significant association between PAH exposure and accelerated methylation ageing. The importance of exposure-related ageing needs to be further investigated. Funding National Key Basic Research and Development Program, Natural National Scientific Foundation, the 111 Project, the Program for Changjiang Scholars, and Innovative Research Team Scholars.

To identify an inbred Chinese pedigree with autosomal recessive muscular dystrophy and analyze the molecular defects.Linkage analysis was conducted using short tandem repeat(STR) markers from the regions associated with limb-girdle muscular dystrophy type 2A(LGMD2A) through 2H. Multi-Western blot was performed with anti-calpain-3, anti-dysferlin, anti-gamma-sarcoglycan, anti-alpha-sarcoglycan, and anti-dystrophin monoclonal antibodies. Mutation was determined by reverse transcriptase-polymerase chain reaction and sequencing.Two-point linkage analysis showed significant Lod scores with markers from chromosome 2p13, the highest two-point Lod scores were obtained with D2S337 (Z(max)=1.86 at theta=0). Multi-Western blot confirmed dysferlin deficiency of muscle specimen from the proband. Mutation analysis revealed a novel 6429delG mutation on exon 53 of the DYSF gene for the proband.The authors identified an inbred Chinese pedigree with Miyoshi myopathy caused by a 6429delG on the DYSF gene. This mutation is predicted to result in premature termination of translation.