Modeling Methods and Influencing Factors for Age Estimation Based on DNA Methylation.

Age estimation based on tissues or body fluids is an important task in forensic science. The changes of DNA methylation status with age have certain rules, which can be used to estimate the age of the individuals. Therefore, it is of great significance to discover specific DNA methylation sites and develop new age estimation models. At present, statistical models for age estimation have been developed based on the rule that DNA methylation status changes with age. The commonly used models include multiple linear regression model, multiple quantile regression model, support vector machine model, artificial neural network model, random forest model, etc. In addition, there are many factors that affect the level of DNA methylation, such as the tissue specificity of methylation. This paper reviews these modeling methods and influencing factors for age estimation based on DNA methylation, with a view to provide reference for the establishment of age estimation models.

Morphological analysis of three-dimensionally reconstructed frontal sinuses from Chinese Han population using computed tomography.

The uniqueness and reliability of frontal sinuses for personal identification have gained wide recognition in forensics. However, few studies have assessed the usefulness of a three-dimensional (3D) model of the frontal sinus for human identification. This study aimed to develop standardized techniques to classify the frontal sinus according to its 3D morphological metrics and discover the usefulness of the 3D frontal sinus model in identification of Chinese Han population. One hundred and ninety-six computed tomography (CT) scans of patients older than 20 years (84 males and 112 females) were collected. A 3D frontal sinus digital model was segmented using Dolphin Imaging software. The following morphological metrics of the 3D frontal sinus were used to develop the coding system: bilateral or unilateral, spatial relationships of the two sides, number of septations, superior volume side, the shape of the 3D model of each side, shape of the medial surface and frontal ostium on each side, number of accessory septations on each side, number of supra-orbital cells of the medial surface and lateral surface on each side, and number of the arcades on each side. The new coding system accurately identified all of our research individuals. This study discovered a number of individual variations in the 3D frontal sinus morphology patterns. A coding system, which is based on these morphological patterns, exposes the morphological variants of frontal sinuses and presents the usefulness of 3D frontal sinus model for human identification.

Computer-aided superimposition of the frontal sinus via 3D reconstruction for comparative forensic identification.

The anatomical uniqueness of the frontal sinus morphology has been widely used for comparative forensic identification using various techniques, mostly including 2D X-rays or one fixed slice of an axial computed tomography (CT) scan image. However, computer-aided 3D automatic graphical comparison techniques can provide accurate comparisons between two 3D models that allow users to comply with even the strictest deviation standards, avoiding error-prone identification of frontal sinuses with similar morphologies. The study proposes the use of a computer-aided comparative paradigm based on the 3D-3D frontal sinus model superimposition process and further assesses the anatomical uniqueness of frontal sinuses using a large Chinese Han sample. Three hundred thirty-six patients older than 20 years with two multi-slice CT scans were collected. Frontal sinus 3D models were semi-automatically segmented through Dolphin Imaging software. Automatic pairwise comparisons of 336 matched pairs from the same person and 340 mismatched pairs from different individuals with an analysis of average root mean square (RMS) point-to-point distance were performed using Geomagic Studio Qualify software. RMS ranged between 0.005 and 1.032 (mean RMS 0.390 ± 0.25 mm) in the group of matches and between 1.107 and 19.363 (mean RMS 4.49 ± 2.69 mm) in the group of mismatches. On average, the RMS value was over ten-fold greater in mismatches than in matches. Statistically significant differences in RMS between the group of matches and mismatches were assessed using the Mann-Whitney U test (p < 0.05). This study supports the value of the frontal sinus with a 3D computer-aided superimposition method for human identification with large samples when DNA, fingerprints, and dental materials are not accessible.

[Method of Identifying Male Lineages Based on Main Haplotype and Analysis of the Distribution of Y-STR Haplotype Mismatch Based on the Bayesian Theory].

OBJECTIVE: To establish a classification method to identify different male lineages in a large population, to study the distribution patterns of Y-STR loci mismatches among Han Chinese male lineage members and to explore the mismatch probability distribution among the members with different meiosis intervals in the family. METHODS: Peripheral blood samples of 269 male individuals from 12 lineages in Han Chinese population and 45 unrelated male individuals were collected. Then, Yfiler Plus TM and ZGWZ FSY or Yfiler Platinum amplification kits were used, obtaining 314 Y-STR haplotypes. The Y-STR haplotype with 3 or more repetitions were selected as the main haplotype, in which the largest number was selected as the first data center. According to the standard of Y-STR genotype, those with mismatches within five loci and six steps were clustered and merged. Then, the main haplotype of the largest number in the remaining data was taken as the second data center, and cluster analysis is carried out in turn until there is no main haplotype remained. Pair comparison was conducted between lineage members and unrelated individuals, and the mismatch distribution among lineage members and unrelated individuals was calculated respectively. The average mismatch rate of each locus was subsequently calculated, as well as the mismatch probability distribution among members with different meiosis intervals within the lineage. RESULTS: 269 out of the 314 individuals were divided into 12 groups by cluster analysis method, accomplishing 100% accuracy between the cluster groups thus identified and the 12 known lineages. The remaining 45 unrelated individuals were scattered. The mismatch loci was within 0-7 loci and 0-7 steps among lineage members and the mismatch between unrelated individuals was at least 11 loci and 15 steps. The mismatch loci with the largest number of one-step and two-step mismatch were different in each lineage and had features that were specific to each lineage. The minimum mutation count and average mismatch rate of each locus were significantly correlated with the mutation rate. Two individuals with no mismatch had a 19.7% probability of 1 meiosis interval and a 71.2% probability of less than 6 meiosis interval. Two individuals with 3 loci mismatches had a 65.2% probability of more than 10 meiosis intervals. CONCLUSION: The cluster analysis method based on main haplotypes provided in this paper can quickly and effectively differentiate large male lineage samples. The clustering method and the mismatch probability distribution of different meiosis intervals obtained thus can provide new ideas for research and screening instruments, and important reference for lineage investigation, data analysis and practical application of Y-STR database in the future.

Application of multiplex methylated-specific PCR with capillary electrophoresis to explore prognostic value of TSGs hypermethylation for hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is a malignant tumor that severely threatens human health. To date, early detection for HCC patients is particularly significant due to their poor survival rates even after liver resection. METHODS: Therefore, an efficient and sensitive detection method for monitoring liver cancer, multiplex methylation-specific PCR (MSP) coupled with capillary electrophoresis, is developed. RESULTS: Simulations demonstrated that the methylation status of RASSF1A, p16, SFRP1, and ELF could be detected even when DNA equaled or exceeded 12.5 ng simultaneously. Also, its accuracy for methylation detection outweighed polyacrylamide gel electrophoresis (87.5%) and agarose electrophoresis (84.3%), reaching 92.1%. Subsequently, we implemented multiplex MSP with capillary electrophoresis to investigate methylation status of the four tumor suppressor genes in tissue specimens and explore the prognostic value for HCC patients. As the data suggested, multivariate cox regression analysis revealed that the recurrence-free survival of 46 patients was greatly associated with portal vein tumor thrombus (PVTT) and p16 methylation and receiver operating characteristic (ROC) curves demonstrated that the predictive range of portal vein tumor thrombus (PVTT) combined with p16 hypermethylation was more sensitive than that of either PVTT or p16 hypermethylation alone with regard to disease recurrence in patients with HCC, which could be testified as a valuable biomarker in Clinical application. CONCLUSION: Multiplex MSP coupled with capillary electrophoresis has an excellent prospect of clinical application for monitoring early liver cancer and screening valuable biomarkers for prognosis of HCC patients.

Engrailed-2 is down-regulated but also ectopically expressed in clear cell renal cell carcinoma.

Engrailed-2 (EN2), a member of the homeobox family of genes, encodes a homeodomain-containing transcription factor that is thought to be a potential oncogene in a number of cancers. Because the role of EN2 in clear cell renal cell carcinoma (CCRCC) has not been determined, we investigated its expression in CCRCC tissues and cell lines. Using immunohistochemical (IHC) staining, we found that EN2 protein was expressed in normal renal cells and tubules, but was frequently down-regulated in tissues from patients with CCRCC and in CCRCC cell lines. In addition, we found that EN2, which functions in the nucleus, was completely localized to the cytoplasm of CCRCC cells as detected by IHC and immunofluorescence staining. Furthermore, expression of EN2 protein was negatively correlated with increasing histological grade of CCRCC tumors (P = 0.003). The exact role of EN2 expression in renal carcinoma carcinogenesis requires further investigation.

A genetic variant in the promoter region of miR-34b/c is associated with a reduced risk of colorectal cancer.

The miR-34 family members, described as potential tumor suppressors, were downregulated in colorectal cancer (CRC). Loss of miR-34 impairs TP53-mediated cell death, while overexpression of miR-34 induces apoptosis. A potentially functional polymorphism (i.e., rs4938723T/C) in the promoter region of pri-miR-34b/c was predicted to influence the GATA-X binding sites. We aimed to investigate the association between miR-34b/c rs4938723 and TP53 Arg72Pro polymorphisms and the risk of CRC. We genotyped the two polymorphisms in 347 CRC patients and 488 healthy controls using polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing assay. We found that the CC genotype and C allele of the miR-34b/c rs4938723 were associated with a significantly decreased risk of CRC compared with the TT genotype and T allele (CC vs. TT: adjusted OR=0.56; 95% CI, 0.34-0.91; C vs. T: adjusted OR=0.78; 95% CI, 0.64-0.97). In combined analysis, a borderline significance was also observed in subjects carrying the rs4938723 CT/CC and TP53 GG genotypes (adjusted OR=0.66; 95% CI, 0.43-0.99). These findings indicate that the rs4938723 in the promoter region of pri-miR-34b/c was a protective factor for the development of CRC. As the significance is marginal, further replication studies are warranted to confirm these results.

[Expression of SSB following early myocardial ischemia in rats].

OBJECTIVE: To investigate the expression of Sjögren's syndrome antigen B (SSB) gene and SSB protein in the early ischemic myocardium in rats. METHODS: Adult healthy Sprague-Dawley rats were randomly divided into groups of operation [myocardial ischemia (MI) and non-ischemia (NI)], non-operation (NO) and sham-operation (SO) (n = 6 for MI and NI; n = 4 for NO and SO). According to time of ischemia, it was then divided into groups of 0 min, 15 min, 30 min, 60 min, 120 min, and 240 min. The expression of SSB gene in the myocardium was examined by real-time PCR, and the expression of SSB protein was examined by Western blot and immunofluorescence staining. RESULTS: The expressions of SSB gene was down-regulated at early stage of ischemia. There was significant difference between 0 min and 120 min at the level of expression of SSB gene in MI group, so did that between 120 min group and NO group (P < 0.05). The expression of SSB protein at 60 min after ischemia was significantly decreased compared with that in the group of 0 min (P < 0.05). The expression of SSB protein in NI groups was significantly higher than that in MI groups at the time of 60 min and 120 min after myocardial ischemia (P < 0.05). Additionally, the expression of SSB protein was mainly located in the myocardial nucleus, myocardial plasma, and plasma membrane of partial myocardiocytes according to the result of immunofluorescence staining. CONCLUSION: SSB may participate in pathophysiologic regulation process in myocardial cells at the early stage of myocardial ischemia in rats.

Interactions of interleukin-12A and interleukin-12B polymorphisms on the risk of intracranial aneurysm.

Several lines of evidence indicate that inflammatory processes play pivotal role in the development of intracranial aneurysm (IA). Recently, polymorphisms in the interleukin-12 (IL-12) gene were shown to be associated with immune-mediated inflammatory disease. The aim of this study was to investigate the interactions of IL-12A and IL-12B polymorphisms on the risk of IA in a Chinese population. A total of 422 individuals (including 164 patients with IA and 258 controls) were involved in the study. The polymorphisms (i.e., rs2243115 and rs568408 in IL-12A and rs3212227 in IL-12B) were genotyped by polymerase chain reaction-restriction fragment length polymorphism assay and DNA sequencing. We found an association of the AC/CC genotypes and C allele of IL-12B rs3212227 with an increased risk of IA, compared with the AA genotype and A allele (AC/CC vs. AA: OR = 2.09, 95 % CI: 1.29-3.38; C vs. A: OR = 1.45, 95 % CI: 1.10-1.91). Moreover, a significant gene interaction of IL-12A and IL-12B was evident on the risk of IA, and subjects carrying variant genotypes of IL-12B rs3212227 had an increased risk of IA. In the stratified analysis by gender, the IL-12B rs3212227 AC/CC genotypes had an increased risk of IA compared with the AA genotype in male patients (AC/CC vs. AA: OR = 4.63, 95 % CI: 1.92-11.16). These findings suggest that the IL-12A and IL-12B independently and jointly be involved in the susceptibility to IA.

The association between two polymorphisms in pre-miRNAs and breast cancer risk: a meta-analysis.

Emerging evidence has shown that miRNAs participate in human carcinogenesis as tumor suppressors or oncogenes. Single nucleotide polymorphism (SNP) which located in the pre-miRNA may affect the processing and then influence the expression of mature miRNA. Previous studies yielded conflicting results as to the association of two common polymorphisms in pre-miRNAs (i.e. hsa-miR-146 rs2910164 and hsa-miR-196a2 rs11614913) with breast cancer. To derive a more precise effect on the association between these polymorphisms and breast cancer risk, we conducted a meta-analysis. Through retrieving PubMed for the period up to May 2010, a total of four studies were identified with 3,007 cases and 3,718 controls for has-miR-146a rs2910164 polymorphism and with 3,287 cases and 4,298 controls for hsa-miR-196a2 rs11614913 polymorphism. We found that individuals carrying CC genotype of has-miR-196a2 rs11614913 polymorphism was associated with an increased breast cancer risk in homozygote comparison (OR = 1.30; 95% CI, 1.01-1.68), and dominant model (OR = 1.11; 95% CI, 1.01-1.23). However, no significant association between has-miR-146a rs2910164 polymorphism and breast cancer risk was observed in all comparison models tested. These findings suggest that has-miR-196a2 rs11614913 polymorphism may play crucial roles in breast cancer development.

RAD51 135G/C polymorphism and breast cancer risk: a meta-analysis from 21 studies.

Growing evidence suggests that RAD51 plays a pivotal role in the repair of DNA double-strand breaks and the maintenance of genomic stability. A single nucleotide polymorphism, 135G/C, has been identified in the 5' untranslated region of the RAD51 gene and has been shown to influence gene transcription activity. Previous studies yielded conflicting results as to the association of 135G/C polymorphism with breast cancer. We aimed to assess the effect of 135G/C of RAD51 on breast cancer susceptibility with the use of a meta-analysis. We performed a meta-analysis of 21 published case-control studies up to April 2010. We found that the CC genotype was associated with a significantly increased risk of breast cancer when compared with the GG, CG, and CG/GG genotypes. Subgroup analyses showed that individuals carrying the CC genotype were associated with an elevated tumor risk in European populations and in sporadic breast cancer. After stratified analyses according to manuscript quality, the CC genotype was associated with a significantly increased risk of breast cancer compared with the CG genotype in studies of both higher and lower quality. However, significantly elevated risk was found in studies of higher quality, but not in studies of lower quality when homozygote and a recessive comparison model were tested. This meta-analysis indicates that RAD51 135G/C polymorphism may be identified as a susceptibility locus for breast cancer.

Characteristics of eight X-STR loci for forensic purposes in the Chinese population.

X-chromosomal short tandem repeats (ChrX STRs) loci are used for forensic practice in recent years. Considering the unique heredity characteristics of ChrX, recombination and linkage disequilibrium (LD) among ChrX STR loci vary between male and female and different populations as well. However, there is a lack of data for analysis of recombination and linkage disequilibrium on ChrX STR loci in the Chinese population. In this work, a total of 303 unrelated individuals (203 males and 100 females) in the Chinese Han population were analyzed with Mentype Argus X-8 PCR amplification kit (DXS10135-DXS8378, DXS7132-DXS10074, HPRTB-DXS10101, and DXS10134-DXS7423). The recombination and linkage disequilibrium of the eight ChrX STR loci were investigated with HapMap LD plots and software ARLEQUIN 3.1. Allele frequencies of the eight loci and further population forensic genetic parameters were obtained. Our results revealed hotspots for recombination, and there was no obvious evidence for LD among the eight loci in the Chinese population. Our work implied that single locus frequencies rather than haplotype frequencies should be applied for forensic practice in the Chinese population.

[Relationship between RNA degradation and postmortem interval in mice].

OBJECTIVE: To investigate the degradation changes of beta-actin mRNA and 18S rRNA in different time points and temperature after death, and to explore the relationship between the changes and postmortem interval (PMI) in the brain of mice. METHODS: Twenty-four health adult C57BL/6 mice were randomly divided into two groups (12 each group). They were sacrificed by cervical dislocation and placed in chamber with two different temperature (4 degrees C and 37 degrees C, humidity was 80%). The mice brains were sampled at 6 different time points(immediately, 0.5h, 2h, 6h, 24h, 48h), and total brain RNA were extracted. Ct value of each sample was obtained using RT-PCR and real-time PCR technology, and beta-actin mRNA and 18S rRNA content ratio was calculated. The correlation between the content ratio and PMI was expressed using statistical regression analysis. RESULTS: At 37 degrees C, RNA degradation rate was faster than 4 degrees C, which showed that there was correlation between temperature and RNA degradation. Comparing with the stability of beta-actin mRNA, 18S rRNA was more stable. CONCLUSION: The study on degradation of beta-actin mRNA and 18S rRNA in mice brain using real time PCR technology could provide a new theoretical basis for estimation of PMI and would be supplementary to the traditional methods.

[Expression of SSB gene in rats with early isehemic myocardium and its implications].

OBJECTIVE: To examine the expression of Sjögren's syndrome antigen B (SSB) gene in rats with early ischemic myocardium. METHODS: Adult healthy Sprague-Dawley rats were randomly divided into four groups (ischemia (MI), non-ischemia (NI), sham-operated (SO) and normal control (NO) groups). The expression of SSB and beta-actin gene in the myocardium was examined by real time PCR at 0 min, 5 min, 15 min, 30 min, 60 min, 120 min after ligation of the left anterior descending coronary artery (LAD). RESULTS: The expression of SSB gene was down-regulated after ischemia. Significantly lower expression of the gene in the rats with myocardial ischemia was found at 120 min after ligation compared with those before ligation and the control group at 120 min after ligation (P < 0.05 ). However, no significant differences were found between other groups (P > 0.05). CONCLUSION: SSB gene may be involved in the pathophysiologic regulating process in myocardial cells at the early stage of myocardial ischemia in rats.

[The expression of Basigin mRNA in early ischemic myocardium and non-ischemic myocardium].

OBJECTIVE: To investigate the expression of Basigin mRNA in early ischemic myocardium (EIM) and non-ischemic myocardium (NIM) in rat. METHODS: Real-time polymerase chain reaction (PCR) technique was applied for detecting Basigin mRNA and beta-actin expression in EIM and NIM at 0 min, 15 min, 30 min, and 60 min. RESULTS: No significant differences of the Basigin mRNA expression in EIM and NIM between 0 min and sham operation (SO) or non-operation group (P > 0.05) were observed. Basigin mRNA expression in EIM significantly decreased in 15 min, 30 min, and 60 min group compared with that of 0 min group, respectively (P < 0.001). However, we failed to find any significant difference in NIM and NO from 0 min to 60 min. Basigin mRNA expression in EIM decreased significantly compared with that of in NIM (P < 0.05) after myocardial ischemia for 30 min. CONCLUSION: Basigin was involved in myocardium pathophysiology process after myocardial ischemia for 30 min, indicating that Basigin may be identified as a predictor of early myocardial ischemia in forensic medicine.

The association of interleukin-16 polymorphisms with IL-16 serum levels and risk of colorectal and gastric cancer.

Interleukin (IL)-16, a multifunctional cytokine, plays a fundamental role in inflammatory diseases, as well as in the development and progression of tumors. Genetic variation in the DNA sequence of the IL-16 gene may lead to altered cytokine production and/or activity, and this variation may modulate an individual's susceptibility to both colorectal cancer (CRC) and gastric cancer (GC). To test this hypothesis, we investigated the association of IL-16 gene polymorphisms with serum levels of IL-16 and the risk of CRC and GC in a Chinese population. We analyzed single-nucleotide polymorphisms of the IL-16 gene in 596 cancer patients (376 patients with CRC and 220 patients with GC), and also in 480 age- and sex-matched controls using polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing methods. Serum IL-16 levels were measured by enzyme-linked immunosorbent assay. The rs11556218 T/G polymorphism of the IL-16 gene was significantly associated with the susceptibility to CRC and GC patients. Both male and female patients carrying the G allele had a significantly higher risk for developing CRC and GC compared with individuals carrying the T allele. Alternatively, women carrying the T allele (rs4072111 C/T) showed a decreased risk for CRC and GC compared with individuals carrying the C allele. In patients with CRC or GC, IL-16 serum levels were significantly higher than those in the healthy controls, although no significant association between IL-16 polymorphisms and serum levels of IL-16 was observed. Our data indicate that IL-16 polymorphisms may contribute to CRC and GC susceptibility.

[Association of interleukin-1B gene polymorphisms with coronary heart disease].

OBJECTIVE: To determine whether polymorphisms in interleukin-1B gene promoter -31 and exon 5 +3953 loci are associated with coronary heart diseases (CHD) in Chengdu Han population. METHODS: Two SNPs of IL-1B gene (+3953C/T and -31T/C) in 100 patients with CHD (CHD group) and 144 healthy controls in Chengdu were analysed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Two genotypes at IL-1B + 3953 locus (CC and CT) and 3 genotypes at IL-1B -31 locus (CC, TC and TT) were identified. The -31T alleles carriers were associated with a significantly increased risk of CHD as compared with the non-carriers (OR = 2.12, 95% CI: 1.45-3.09, P < 0.01). Genotypes and allele frequencies at IL-1B + 3953 locus in CHD cases did not differ from the controls. CONCLUSION: IL-1B -31T/C polymorphism may contribute to the risk of developing CHD in Chengdu Han population.

[Genetic polymorphism of NFKB1 gene in Chinese Han of Chengdu and Thai populations].

OBJECTIVE: To investigate the single nucleotide polymorphisms (rs3774963 C>G and rs11722146 A>G) of NFKB1 gene between the Chinese Han of Chengdu and Thai populations, and simultaneously to compare distributions of genotype and allelic frequencies of NFKB1 among different ethnic groups from the International Haplotype Map Project. METHODS: The genotypes and allele frequencies of NFKB1 gene rs3774963 C>G and rs11722146 A>G were analyzed in 118 healthy Chinese Han of Chengdu and 101 Thai individuals using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) strategy and DNA sequencing. RESULTS: The frequencies of the CC, CG, and GG genotypes of rs3774963 C>G were 15.3%, 43.2%, and 41.5% in Chinese Han of Chengdu, and 25.7%, 47.5%, and 26.7% in Thai population, respectively. The frequencies of the C and G alleles were 36.9% and 63.1% in Chinese Han of Chengdu, and 49.5% and 50.5% in Thai population, respectively. There were significant differences in the genotypes and allele frequencies between the two groups. The frequencies of the AA, AG, and GG genotypes of rs11722146 A>G were 22.9%, 50.0%, and 27.1% in Chinese Han of Chengdu, and 18.8%, 53.5%, and 27.7% in Thai population, respectively. The frequencies of the A and G alleles were 47.9%, 52.1% in Chinese Hen of Chengdu, and 54.5%, 45.5% in Thai population, respectively. However, no statistically significant difference was observed between the two populations. Interestingly, when compared with the data from the International Haplotype Map Project, the genotypes and allele frequencies of NFKB1 gene rs11722146 A>G but not rs3774963 C>G in the Chinese Han of Chengdu were significantly different from those among European and Sub-Saharan African populations. CONCLUSION: NFKB1 gene polymorphism in diverse populations is significantly different.

Circular Ribonucleic Acid Expression Profile in Mouse Cortex after Traumatic Brain Injury.

Traumatic brain injury (TBI) causes high rates of worldwide death and morbidity because of the complex secondary injury cascade. Circular ribonucleic acid (RNA) (circRNA), a type of RNA that forms a covalently closed continuous loop, may be involved in the regulation of secondary injury because it is expressed widely in the brain and contributes to a large class of post-transcriptional regulators. Deep RNA sequencing (RNA-seq) and bioinformatic analysis were performed to investigate the expression profile and function of circRNAs in the mouse cortex after controlled cortical impact (CCI). A total of 19,794 circRNAs were identified, and 1315 were annotated in circBase. There were 191 filtered differentially expressed circRNAs (98 for up-regulated and 93 for down-regulated). The gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses indicated that inflammation, cell death, and repair of damage were the main biological processes and molecular mechanisms related to altered circRNAs. The pathway-circRNA interaction network revealed three core circRNAs and five corepathways related to TBI. The circRNA-messenger RNA (mRNA) interaction network and competitive endogenous RNA (ceRNA) analysis suggested potential microRNA (miRNA) sponges and target mRNAs. In addition to five optimal circRNA-miRNA-mRNA pairs were analyzed, circRNA_16895-miRNA myosin-10 (Myo 10) was predicted to regulate fragment crystallizable gamma receptors (FcγR)-mediated phagocytosis pathway. Four circRNAs were selected for quantitative real-time polymerase chain reaction analysis to validate the sequencing data. Our results provide promising functions of circRNAs aberrantly expressed in TBI to explore molecular mechanisms and potential therapeutic targets for its therapy.

R497K polymorphism in epidermal growth factor receptor gene is associated with the risk of acute coronary syndrome.

BACKGROUND: Previous studies suggested that genetic polymorphisms in the epidermal growth factor receptor (EGFR) gene had been implicated in the susceptibility to some tumors and inflammatory diseases. EGFR has been recently implicated in vascular pathophysiological processes associated with excessive remodeling and atherosclerosis. Acute coronary syndrome (ACS) is a clinical manifestation of preceding atherosclerosis. Our purpose was to investigate the association of the EGFR polymorphism with the risk of ACS. In this context, we analyzed the HER-1 R497K and EGFR intron 1 (CA)n repeat polymorphisms in 191 patients with ACS and 210 age- and sex-matched controls in a Chinese population, using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) strategy and direct sequencing. RESULTS: There were significant differences in the genotype and allele distribution of R497K polymorphism of the EGFR gene between cases and controls. The Lys allele had a significantly increased risk of ACS compared with the Arg allele (adjusted OR = 1.49, 95% CI: 1.12-1.98, adjusted P = 0.006). However, no significant relationship between the number of (CA)n repeats of EGFR intron 1 (both alleles < 20 or any allele > or = 20) and the risk of ACS was observed (adjusted OR = 0.97, 95% CI: 0.58-1.64, adjusted P = 0.911). Considering these two polymorphisms together, there was no statistically significant difference between the two groups. CONCLUSION: R497K polymorphism of the EGFR gene is significantly associated with the risk of ACS. Our data suggests that R497K polymorphism may be used as a genetic susceptibility marker of the ACS.