The association of the aldehyde dehydrogenases-2 (ALDH2) Glu504Lys polymorphism  and cancers has been investigated. This meta-analysis aims to comprehensively assess the influence of this polymorphism on the overall cancer risk.Eligible publications were retrieved according to inclusion/exclusion criteria and the data were analyzed using the Review Manager software (V5.2).P = 0.02, 2I = 92%). Subgroup analysis by cancer type indicated a significantly increased UADT cancer risk  in individuals with the Lys+ genotypes. Subgroup analysis by country indicated that individuals from Japan with the Lys+ genotypes had a significant 38% increased cancer risk .A meta-analysis based on 51 case-control studies consisting of 16774 cases and 32060 controls was performed to evaluate the association between the ALDH2 Glu504Lys polymorphism and cancer risk. The comparison of genotypes Lys+ (Lys/Lys and Lys/Glu) with Glu/Glu yielded a significant 20% increased cancer risk (OR = 1.20, 95%CI: 1.03–1.39, Our results indicated that the ALDH2 Glu504Lys polymorphism is a susceptible loci associated with overall cancers, especially esophageal cancer and among Japanese population. Based on available epidemiological data, alcohol ingestion is shown to be carcinogenic to humans and causally related with liver, colorectal, female breast and upper aerodigestive tract (UADT) cancers . ApproxiAldehyde dehydrogenase-2 (ALDH2) is expressed in the liver as well as gastrointestinal tract. It belongs to a low-Km mitochondrial ALDH and is the second enzyme to eliminate most of the acetaldehyde generated during alcohol metabolism in vivo . Human ATherefore, it is hypothesized that the genetic polymorphism in ALDH2 gene may be strongly correlated with the susceptibility to cancer, and a number of studies have investigated the association between ALDH2 Glu504Lys polymorphism and cancer risk. Most of the studies focused on esophageal cancer, followed by colorectal cancer, head and neck cancer, etc. In a meta-analysis by Yang et al , ALDH2 5Publications were searched via the PubMed bibliographical database with the last update as of 30 April, 2014. The following keywords and MeSH terms were used: [“aldehyde dehydrogenase 2” or “ALDH2”] and [“polymorphism” or “genetic polymorphism” or “mutation” or “variation” or ”variant” or “single nucleotide polymorphism” or “SNP”] and [“cancer” or “malignant tumor” or “malignant neoplasm”]. As a prerequisite, only studies published in English were identified. All eligible studies were retrieved and the full text of the articles was examined to make sure the data of interest were included. In addition, if multiple reports from the same patients were found, only the publication with the most complete data set was included. If more than one ethnic population or cancer type were included in one article, data were extracted separately for each ethnic population or cancer type whenever possible.Studies that we identified were required to meet the following criteria: (1) study on the evaluation of the ALDH2 Glu504Lys polymorphism and cancer risk; (2) case-control study that used either population- or hospital-based designs; (3) study that contained complete information about all genotype frequency. Studies were excluded if they were case-only studies, review articles, or reports without usable data.Two investigators (QC and JW) independently extracted the following information from all selected articles: first author, year of publication, country of origin, population ethnicity, study design , cancer type, genotyping information . Ethnic backgrounds were categorized as Asian, African or Mixed (composed of different ethnic groups). Cancers of oral cavity, oropharynx, hypopharynx, larynx, esophagus and stomach were defined as upper aerodigestive tract (UADT) cancer .χ2 test (P>0.05)   . When thwas used ; otherwiwas used . Sensitifs) with the significance set at 0.05 for each meta-comparison. If the calculated Nfs value was smaller than the number of observed studies, the result might run the risk of having publication bias. We calculated the Nfs0.05 according to the formula Nfs0.05 = (ΣZ/1.64)2-k, where k is the number of included articles [Publication bias was evaluated by using the fail-safe number  and genetic model comparison . A random effects model was performed in these analyses. Then we performed subgroup analysis based on cancer type, country, study design and sample size and assessed the source of heterogeneity for genetic model comparison (Lys+ vs Glu/Glu). As a result, cancer type  and country  but not study design  or sample size  were the significant sources of heterogeneity.In the pooled analysis, we have found heterogeneities in allelic model comparison  and subjects <300 . Moreover, the estimated pooled odds ratio in other subgroups did not change, which suggested that the results of stratified analyses were also stable.Influence analysis was conducted by repeating the meta-analysis while excluding the studies that were not in HWE or the fs) at a significance level of 0.05 for each comparison. The Nfs0.05 values for the comparison of Lys versus Glu (Nfs0.05 = 2767), Lys+ versus Glu/Glu (Nfs0.05 = 4998) were greater than the number of studies included in the meta-analysis.Lastly, to assess publication bias, we calculated the fail-safe number  appear to be associated with an increased risk of cancer. Due to the prevalent of ALDH2 polymorphism in approximately half of East Asians but absent in Europeans and Africans  and possPHWE values were not available, interestingly, we found the effect of variant allele (504Lys) on colorectal cancer was contrary to that on UADT cancer. Recently, Zhao et al [In the subgroup analysis by cancer type, significantly increased risk of UADT cancer with ALDH2 polymorphism was observed but no significant association was found among studies of other cancers . In the UADT cancer subgroup, we further performed position-specific analyses and the results showed that individuals with the variant allele (504Lys) significantly increased 52% risk of esophageal cancer, 22% risk of head and neck cancer and 18% risk of gastric cancer. Some previous meta-analysis had reported the similarly elevated risks –74. Furtao et al  had repoIn the general population, the variant 504Lys allele is prevalent in Northeast Asian individuals  . After sPHWE values were not available. Except the cancer-specific analysis of colorectal cancer group and size-specific analysis of subject <300 group, the results were persistent and robust, suggesting that this factor had little effect on the overall estimates.Heterogeneity is a potential problem when interpreting the results of a meta-analysis, and identifying the sources of heterogeneity is one of the most important goals of meta-analysis. In the present study, significant between-study heterogeneity in the pooled analyses of all included studies was found in both allelic and genetic models. To find the sources of heterogeneity, we performed subgroup analyses stratified by cancer type, country, study design and sample size. Our results indicated that the sources of heterogeneity were from cancer type and country, suggesting that the results of cancer-specific and country-specific analysis were reliable. Furthermore, if the distribution of genotypes in the control groups were not in HWE, the results of the genetic association studies might be spurious. Hence, we performed sensitivity analysis by excluding the studies that were not in HWE or the Despite the clear strength of our study including large sample sizes, some limitations of this meta-analysis should be mentioned. First, since the negative findings are usually difficult to get published or only published in some non-English journals, the ones that reported in other languages may bias the present results. Second, the present study was based on unadjusted ORs, and the confounding factors such as age may still bring some bias. Third, as the lack of sufficient original data, we could not conduct subgroup analysis according to drinking status that may influence the cancer risk. Forth, besides ALDH, activity of alcohol dehydrogenase (ADH) that is responsible for oxidation of ethanol to acetaldehyde can also play an important in the accumulation of acetaldehyde ; therefoIn conclusion, this meta-analysis indicated that the Glu504Lys polymorphism of ALDH2 gene is a candidate for susceptibility to overall cancers, especially in esophageal cancer and among Japanese population. Moreover, due to the limitations mentioned above, well-designed studies taking into consideration of gene-gene and gene-environment interactions should be performed to confirm such associations.S1 Table(XLSX)Click here for additional data file.S1 Checklist(DOCX)Click here for additional data file.S2 Checklist(DOC)Click here for additional data file.


MCP-1 -2518 polymorphism and pulmonary tuberculosis. Our new classification considers genetic distance, migration and linguistic origins, which will increase homogeneity within ethnic groups.In meta-analyses of genetic association studies, ancestry and ethnicity are not accurately investigated. Ethnicity is usually classified using conventional race/ethnic categories or continental groupings even though they could introduce bias increasing heterogeneity between and within studies; thus decreasing the external validity of the results. In this study, we performed a meta-analysis using a novel ethnic classification system to test the association between We included thirteen studies from three continents  and considered seven ethnic groups .MCP-1 -2518 polymorphism and TB susceptibility only in the East Asian and Latin American groups . This association is not observed in other ethnic groups that are usually considered in the Asian group, such as India and Persia, or in the African group.The results were compared to the continental group classification. We found a significant association between MCP-1 -2518 polymorphism and TB susceptibility only in the East Asian and Latin American groups. We suggest the use of our new ethnic classification in future meta-analysis of genetic association studies when ancestry markers are not available. This new classification increases homogeneity for certain ethnic groups compared to the continental classification. We recommend considering previous data about migration, linguistics and genetic distance when classifying ethnicity in further studies.There is an association between  MCP-1 belongs to a group of CC chemokines located in chromosome 17q11.2. MCP-1 protein interacts with chemokine C-C motif receptor 2 (CCR2) to activate and recruit monocytes, macrophages, CD4+ T cells and immature dendritic cells to the site of infection ) AND "Tuberculosis"[Mesh]. Mesh term “MCP-1” gave the result CCL2. “Reference sequence 1024611 A/G” gave zero results. Our selection criteria included: 1) studies evaluating the association between MCP-1-2518 A/G and TB risk, 2) observational studies, 3) pulmonary TB, 4) studies performed in adults and children, 4) patients without HIV or cancer, 5) available allelic and the genotype frequencies to estimate an odds ratio (OR), 6) control groups that met Hardy Weinberg Equilibrium, and 7) articles published until December 2013. Studies that did not meet these criteria were excluded. When original articles included more than one study population, we considered each as an independent study. In case of multiple publications on the same study, we included the study with the larger sample and/or the most recently published. The data search retrieved 23 articles. Ten studies were excluded because they were reviews or meta-analyses, or corresponded to pediatric populations, spinal TB, latent TB, HIV positive individuals or data from controls was inaccessible. At the end, 13 studies (7651 cases and 8056 controls)  All articles were separately extracted, reviewed and collated by two independent reviewers who checked for any discordance and reached a consensus in all items. Authors were contacted by email when we needed more information about an article. The following information was extracted for each study: author, year of publication, country of origin, ethnicity, sample size, type of study population, TB definition, allele and the genotype frequencies in cases, controls and methods. The information was systematically reviewed using STROBE and STREGA parameters , 32.We proposed a new ethnic classification based on previous information about genetic distance, migration and linguistic origins , 33–35 aX2 statistic. Genotypes deviated from HWE if two-sided p values were <0.05. Begg funnel plot and Egger’s test indicated publication bias if p value was <0.05. Sensitivity analysis was performed by removing one study at a time to assess the stability of the meta-analysis results.For each study, the Hardy Weinberg Equilibrium (HWE) was calculated for the controls using χ2 based Q test and I2 statistic. P values less than 0.01 were considered significant for heterogeneity. To assess the magnitude of association (pooled OR), in the presence of homogeneity, we used a fixed effects model (inverse variance weighted). Otherwise, we used a random effects model . Pooled OR for the association between MCP-1 - 2518 A/G polymorphism and pTB risk was determined in three steps. First, we did an allelic comparison (G vs. A) to determine the pooled OR in the overall data and by ethnic subgroups. Second, using our new ethnic classification, we analyzed four genotype models: a) recessive (GG vs. AG + AA), b) homogenous co-dominant (GG vs. AA), c) heterogeneous co-dominant GA vs. AA) and c) dominant (GG + GA vs. AA). Third, we compared these results to those obtained from the analysis using the conventional classification. Odds ratio estimates were considered significant if P was <0.05, and were expressed using a 95 % confidence interval (CI). When analyzing by ethnicity, we used the groups that had ≥1 degree of freedom. For our analysis, the wild type allele was A, and the risk allele was G. We did not adjust our model for environmental effects. The statistical analysis was performed using STATA 11.0. .To prove our hypothesis, we assessed heterogeneity and the magnitude of association for each ethnic group. We assessed heterogeneity by using the P = 0.1; Fig. The conventional ethnic analysis found heterogeneity in the three continental study groups for both allelic and genotype analysis. Using our new classification, we found homogeneity for South Asia (India) and West Africa, which were further analyzed with a fixed effects model. We could not improve homogeneity within the rest of the ethnic groups and used a random effects model for their analysis. According to the Begg’s funnel plot and Egger’s test, we did not find any bias in the analysis of the entire group . Interestingly, our new ethnic classification showed that in the Asian continent, the G allele increases risk only in the East Asian ethnic group , but not for South Asian and Persia  but not in Africa (22 % in cases vs. 20 % in controls). Our new ethnic classification showed that East Asia has the highest frequency of the G allele (57 % cases and 46 % controls) in the Asian group. In Latin America, this allele has a similar frequency in Mexico and Peru. In contrast, the African ethnic groups  have a low frequency of G allele in a similar proportion . The recessive model also showed increased susceptibility in both continents but to a lesser extent . Our new ethnic classification showed similar results for Latin America but not for Asian ethnic groups. Only the East Asian group that had the MCP-1 -2518 polymorphism in a homozygote co-dominant and recessive model had an increased risk to develop pTB . The new classification did not find any association for the Persian and South Asian groups. Neither the new nor the continental group classifications found any association in Africa or any of its ethnic groups . The new ethnic classification showed homogeneity for West Africa and South Asia . We found heterogeneity within Arabia, East Asia and Latin America . We could not obtain results for South Africa and Persia, since there was only one study in each group.We found heterogeneity within the ethnic groups from the conventional classification  with countries sharing similar characteristics. This new classification must be further evaluated with new studies related to genetic susceptibility for infectious and noninfectious diseases.Regarding TB susceptibility, our new classification, in contrast to the conventional classification, helped to clarify that the association between MCP-1 -2518 A/G polymorphism and pTB is specific for certain populations such as East Asia and Latin America. To our knowledge, this is the first meta-analysis that uses a model of genetic susceptibility for pTB to assess if a new ethnic classification based on previous findings about genetic distance, migration and linguistic origins, improves homogeneity within each ethnic groups . Thus, wMCP-1- 2518 A/G polymorphism and pTB, which is significant for Asia and South America [MCP-1 -2518 G allele and pTB. Failure to reach homogeneity could be explained because of gene-gene or gene-environmental interactions. It has been reported that people carrying polymorphism MCP-1 -2518 and MMP-1 -1607 have a higher risk to develop severe TB [Previous meta-analyses that use a continental group classification found an association between the  America , 14. Howevere TB . The higevere TB .Recent studies from Africa show that the G polymorphism is not common among this population , 24. HumMCP-1 -2518 G polymorphism. In contrast to South Asia, we found an association between this polymorphism and pTB in East Asia where we also found heterogeneity. Interestingly, India (South Asia) and China (East Asia) accounted together for more than 40 % of TB cases worldwide in the last decade [To deal with heterogeneity in Asia, we considered three groups: East Asia, South Asia and the Middle East –39. The t decade . Howevert decade . Thus, et decade , 49 becat decade .MCP-1 -2518 G allele and other genetic markers between East Asians and Latin Americans. Since Mexico and Peru share migration, common history, language routes and admixture indexes [Latin American ethnic groups that originated from a Han Chinese migration to South America 3000 years ago share HLA markers with this population . This si indexes , 51–53, The limitations in our study are very common to meta-analyses about genetic association studies. We could not consider environmental and genetic factors that influence this association because this information was not found in the original articles. Thus, for research in multifactorial diseases such as TB, we strongly recommend future studies to include information about malnourishment, socioeconomic factors, BCG and TST status, which could also help to control for heterogeneity.MCP-1 -2518 polymorphism is associated to susceptibility. However, despite tuberculosis susceptibility found in Latin America and East Asia due to MCP-1 -2518 polymorphism, the populations within each group are still genetically different.We obtained homogeneity within South Asia and West Africa, where we can rule out that Genetic association studies in populations from Persia, South Africa, South Asia, East Asia and the Americas, where infectious diseases represent a public health problem, will help assess heterogeneity in order to understand the role of ethnicity in genetic susceptibility to these diseases. In the absence of an adequate classification that groups similar genetic characteristics, suitable for understanding genetic susceptibility, our new classification might be a potential proxy for ethnic classification in meta-analysis of genetic association studies when genetic markers are not available.MCP-1 -2518 polymorphism and pTB susceptibility, specifically in Latin American and East Asian populations not detected by using conventional classification. We encourage the use of our new ethnic classification in further genetic association studies for infectious and non-infectious diseases.In summary, using this novel approach, we found an association between the


Ptrend < 0.001 and Ptrend = 0.003, resp.). However, no association remained significant after Bonferroni correction under any model of inheritance. Our results suggest no association between rs2910164 polymorphism with PTC and patients' clinicopathological characteristics.The incidence rate of papillary thyroid cancer (PTC) has increased over the past decades, but the pathogenesis remains unclear. rs2910164, located in pre-miR-146a, has been studied in PTCs with different ethnicity, but the results were inconsistent. Here we evaluate the association between rs2910164 polymorphism and PTC and investigate the effect of this polymorphism on patients' clinicopathological characteristics. 1238 PTC patients and 1275 controls, all Han population, from Northern China, were included in our study. rs2910164 was genotyped using Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS). Analysis of inheritance model was performed using the SNPStats program. Strength of association was assessed by odds ratio (OR) and 95% confidence interval (CI). Overall, no statistical difference in rs2910164 genotype distribution and allelic frequencies between cases and controls was found, and patients with different genotypes had similar clinicopathological characteristics in terms of stage, location, concurrent of benign thyroid tumor, and thyroiditis, while, as the number of G alleles increased, proportion of patients aged ≥45 years and those without metastasis increased ( Thyroid cancer is the most common endocrine malignancy with an incidence rate that has doubled over the past ten years and ranked fifth in 2014 6%) from a previous eighth position 3%) as a cause of all new cancer cases in females % as a ca. Concern% from a MicroRNAs (miRs) are noncoding RNAs that are transcribed from endogenous DNA molecules. The first miR was identified in 1993 and more than one thousand miRs have been discovered in human genome by now . Most miPre-miR-146a C/G polymorphism, designated rs2910164, is encoded on chromosome 5q33 and located in the precursor stem region, +60 relative to the first nucleotide of pre-miR-146a, opposite to the mature miR-146a sequence . It is tWith a case-control study design, 1238 PTC patients and 1275 healthy controls were involved in our study. All PTC patients were pathologically diagnosed and received surgical treatment at the China-Japan Union Hospital of Jilin University in Changchun, Jilin province, China, from January 2010 to December 2014. The histological type of the PTCs was classical PTC, not the follicular variant PTC (fvPTC). The controls consisted of two groups: 360 participants recruited from the First Bethune Hospital of Jilin University in Changchun, Jilin province, China, for a routine health examination, as previously published , and 915For DNA extraction, about 5 mL peripheral venous blood was collected from each subject and stored at −20°C in nonanticoagulant, plexiglass tubes with the ClotBlood DNA kit  used for genomic DNA extraction and ultraviolet spectrophotometer  used for measuring the concentration and purity of the extracted DNA.For SNP genotyping, we used the Assay Designer 3.1 to design the primers of polymerase chain reaction (PCR): 5′-ACGTTGGATGCCACGATGACAGAGATATCC-3′ (forward) and 5′-ACGTTGGATGGAACTGAATTCCATGGGTTG-3′ (reverse). PCR was performed as previously published . The gent-test between groups. Chi-square test was used to evaluate the departure from Hardy-Weinberg equilibrium (HWE) in controls and to compare the difference between cases/controls or subgroups of categorical variables. Analysis of inheritance model was performed using the online SNPStats program (http://bioinfo.iconcologia.net/SNPStats) [P value less than 0.05 was considered statistically significant.Continuous variables were presented with mean and standard deviation and tested using independent samples NPStats) . StrengtP > 0.05), and our study subjects were balanced. Among the cases, 90.6% and 86.4% had no prior self or family history of thyroid disease, respectively, and 11.2% had ever smoked. A majority of patients were at an early stage of disease and 33.6% had lymph node metastasis. Cancer tissues were mostly located at unilateral thyroid (70.5%) and more than one-fourth were bilateral. About half (50.9%) of all patients were diagnosed with thyroid microcarcinoma (a subtype of thyroid cancer with tumor diameter less than 1.0 cm). Besides, 611 (51.5%) patients had benign thyroid cancer(s) simultaneously, including 582 nodular goiter, 35 simple goiter, and 74 adenoma. Among them, 167 patients (39.3%) had their benign tumor on the same side of thyroid gland as the PTC. Moreover, 108 patients had thyroiditis, accounting for 8.7%. The characteristics of the study subjects are shown in Our study included 1238 PTC cases and 1275 healthy controls. The mean ages of cases and controls were 43.43 and 43.38 years, respectively, and the female-to-male rate ratio was more than 2.5 in both groups. Distributions of age and gender were similar between the two groups (P = 0.628). No statistically significant difference in rs2910164 genotype distribution and allelic frequencies was observed between cases and controls .HWE test indicated that the genotype distribution in control group was consistent with that expected under the HWE . To further estimate the effect of rs2910164 polymorphism on PTC clinicopathological characteristics, we performed the inheritance model analysis adjusted by age and gender. In general, codominant, dominant, recessive, overdominant, and log-additive models were conducted to compare the clinical characteristics among patients with different genotypes (P > 0.05), while, compared with CC homozygotes, patients with CG genotype were more likely to have smaller tumor size under the codominant model , and similar result was found for CG/GG carriers under the dominant model . However, no association remained statistically significant after Bonferroni correction. Therefore, we speculated that no association between rs2910164 polymorphism and PTC or clinicopathological characteristics existed.enotypes . For staScientific research on gene polymorphisms involved in carcinogenesis and cancer progression has been a hot area of interest for several decades and the advent of the next generation sequencing technology and GWAS is accelerating the pace of development in this area. Many studies have revealed the importance of miRs in various biological processes, including their roles in several cancers , and provided us with useful information –30.P < 0.001), and heterozygote CG was found to exert a significant influence on PTC occurrence. Thus, we speculate that ethnic diversity may have an influence on the effect of rs2910164 on PTC. Two other studies on participants in Europe  had similar genotype distribution with that of Jazdzewski et al. but found no association between rs2910164 and PTC [Pre-miR-146a was first found in mouse and several studies have shown that it plays an important role in tumorigenesis and cancer metastasis –34. The  and PTC . One of In some retrospective case-control studies carried out on Chinese Han ethnic subjects, rs2910164 has been found to be associated with the risk of several kinds of cancers. Zhou et al. found that the GG genotype of rs2910164 was significantly associated with an increased risk of gastric cancer, which was more notable in the pooled analysis when another follow-up replication study was added, and the association was more significant in younger individuals than the older ones . This fiIt should be mentioned that several meta-analyses tried to explain the association between rs2910164 and cancer risk but got no significant results. Two studies reviewed genetic association studies on common microRNA polymorphisms and hepatocellular carcinoma susceptibility, but no significant association was found for rs2910164 , 41. MorAge, as a well-recognized prognostic determinant in well-differentiated thyroid cancer (WDTC), has been adopted in many thyroid cancer predictive models and prognostic scoring systems –48. AccoOur study demonstrated no association between rs2910164 polymorphism and risk of PTC, as well as its clinicopathological characteristics, in a Chinese Han population. Our results should be interpreted with caution for the ethnicity restriction and relatively small sample size. Furthermore, a considerable number of controls who self-reported no metabolic disease, thyroid disease, or malignancy were included in the study, which might be a potential for bias in the study. However, it is necessary to conduct that further studies with larger sample size are needed to validate our results, and studies with more detailed data estimating the interaction between rs2910164 and environmental factors may provide a better understanding of the role of rs2910164 in PTC development. Nevertheless, our study provides evidence for further studies on different ethnicity and larger sample size in this field.


MicroRNAs (miRNAs) are short, non-coding RNAs that negatively regulate target genes. A single nucleotide polymorphism (SNP) in a miRNA sequence may alter miRNA expression and/or maturation, which was proposed to associate with the development and progression of cancer. The rs895819 polymorphism, located in the terminal loop of pre-miR-27a, has been reported to have relevance to several cancers. In this study, we investigated the possibility of association between polymorphism in rs895819 and susceptibility to colorectal cancer (CRC).We identified a single SNP, rs895819 in pre-miR-27a, for further investigation, were determined in 205 CRC patients and 455 healthy controls.When taking the AA genotype as a reference, we found that AG genotype was not statistically significantly associated with the risk of CRC . However, the GG genotype was significantly associated with risk of CRC . In the AG + GG vs GG group, no significant difference was detected . GG genotype and G allele was associated with an increased risk of metastasis in this study .This study found significant association between rs895819 polymorphism in pre-miR-27a and CRC risk. Population-based studies with large number of subjects and long-term follow-up are needed to verify the association of miR-27a polymorphism with CRC susceptibility and severity.http://www.diagnosticpathology.diagnomx.eu/vs/2061490734125077The virtual slide(s) for this article can be found here:  Colorectal cancer (CRC) is reported to be the third most common cancer worldwide with an estimated one million new cases and it is one of the most frequent cancers and a common cause of cancer-related deaths –3. ImproCRC development involves a multi-step process including both genetic and epigenetic changes, which leads to activation of oncogenes and inactivation of tumor suppressor genes in cancer cells . MicroRNThe relationship between miR-27a and several different cancers have been discussed. Tang et al. reported that the expression of miR-27a was significantly correlated with clinic pathological parameters, including tumor size, lymph node metastasis and distant metastasis, but not with receptor status . PatientWe utilized public databases to identify deregulated miRNAs in CRC and SNPs in these miRNAs sequences, including primary, precursor and mature miRNAs. Our searches identified a single SNP, rs895819 in pre-miR-27a, for further investigation. The rs895819 polymorphism is located in the terminal loop of pre-miR-27a and involves an AG nucleotide transition. The association between rs895819 polymorphism and risk of several cancers were reported –14. In tThis study was approved by the institutional review board of Qilu Hospital of Shandong University. All the participants were voluntary and provided written informed consent prior to taking part in this research.All subjects were recruited from Qilu Hospital of Shandong University between January 2010 and December 2013. They were all newly diagnosed, histopathologically confirmed and without a prior history of cancer or previous chemo- or radiotherapy. In all, 205 patients with CRC were recruited, all of whom were unrelated ethnic Han Chinese population. A structured questionnaire on demographics and environmental exposure, including age, sex, smoking consumption and alcohol drinking, was conducted by trained interviewers through face-to-face interviews with the patients. In addition, 5 ml venous blood was collected from each patient for genomic DNA extraction. The clinical characters of all cases were recorded by reviewing the databases or counseling the doctor.For both the cases and control cohorts, 1.5 ml of whole blood was extracted from each participant and stored at −80°C before being used. DNA from each sample of whole blood was extracted with the QIAamp DNA mini Kit , as directed by the manufacturer's instructions. The concentration of DNA and the purity of each sample were measured by an ultraviolet spectrophotometer . DNA samples were routinely stored at − 20°C.Genotyping was performed by TaqMan allelic discrimination assays using an ABI 7900 system . The prime and probes are as follows: forward, GGCGGAACTTAGCCACTGT, reverse, CAGGGCTTAGCTGCTTGTG. The TaqMan assays were performed in a final reaction volume of 5 ml containing 0.25 ml primer, 0.125 ml probe, 2 ml PCR mixture reagent and 25 ng DNA. The PCR reaction consisted of an initial step at 95 uC for 10 min followed by 55 cycles of denaturing at 95 uC for 15 s and annealing at 60 uC for 60 s. SDS allelic discrimination software (ABI) was used to analyze the PCR genotyping results. Two blank (water) controls were included in each 384-well assay. At least 10% of samples were randomly selected for repeat analysis, yielding 100% concordance. A further 60 samples were selected randomly for direct sequencing to confirm the TaqMan results. Again, the results showed 100% concordance.2 -test with one degree of freedom. P < 0.05 was considered representative of a departure from HWE. Comparison of allele type and genotype distributions in the patients and healthy controls were performed by means of two-sided contingency tables using the χ2 -test. The odds ratio (OR) and 95% confidence interval (CI) for genotypic-specific risk were calculated using an unconditional logistic regression model. P < 0.05 was considered significant for all statistical analyses.Statistical analysis was performed using the SPSS 20.0 software package . Hardy–Weinberg equilibrium analysis was performed using a goodness-of-fit χA total of 205 CRC patients and 455 healthy controls were enrolled in our study. All of the subjects were ethnic Han Chinese. Demographic and other selected characteristics of cases and controls were presented in Table With regard to clinic pathologic characteristics of CRC patients, most patients had a smaller tumor size . Well-moderately status was in 126 (61.46%) patients, while poorly-undifferentiated status was in 79 (38.54%) patients. A total of 86 patients (41.95%) had metastasis among all the patients. According to the TNM classification criteria, 138 and 67 patients were in stage I-II and III-IV, respectively . Taken together, these data suggest that the miR-27a binding site polymorphism rs895819 may be associated with the risk of CRC.In the stratified studies, we attended to detect the association between rs895819 polymorphism and clinicopathological characteristics of patients with CRC. We observed no association between rs895819 polymorphism and the progression of CRC including tumor size and tumor stage. However, GG genotype and G allele was associated with an increased risk of metastasis in this study  Table .Table 4According to the polygenic model of carcinogenesis, unfavorable combinations of polymorphic genetic variants in low-penetrance susceptibility genes contribute to increasing CRC risk. Studies have shown that profiles of miRNA expression differ between normal and tumor tissues, which vary among tumor types and the down-regulation of miRNA subsets implies a tumor-suppressor function, which is often observed in tumor development. Recently, a class of novel functional polymorphisms in miRNAs or their binding sites has attracted great interest. One important SNP in pre-mir-27a of an A to G change was identified as rs895819, and its role in cancer susceptibility has been discussed recently , 15. ThoA previous study by Fu et al. identified 32 differentially expressed miRNAs and 2916 mRNAs from CRC samples and their corresponding normal epithelial tissues by miRNA and mRNA microarray, respectively. Additionally, 6 up-regulated miRNAs  and 4 down-regulated predicted target mRNAs  were selected to validate the expression level and their anti-correlationship in an extended cohort of CRC patients by qRT-PCR . It coulSingle nucleotide polymorphism of miRNA would influence the expression of miRNA and thus would influence the cancer incidence or progression. Zhou et al.  reportedThe association between miR-27a SNP and CRC risk was also studied in several studies. Hezova R et al.  conducteOur results showed an association between miR-27a polymorphism and CRC in Chinese Han patients. Besides, miR-27a polymorphism showed a significant association with the risk of metastasis. However, no functional validation of these observational findings was conducted and a series of following studies would be conducted. Considering that this is from a case-control study, however, population-based studies with large number of subjects and long-term follow-up are needed to verify the association of miR-27a polymorphism with CRC susceptibility and severity.Zaiqiu Wang and Xiaoli Sun are co-first authors.


XRCC1 codon 399 polymorphism and PCA risk has been extensively reported, the results have been ambiguous.The polymorphism in codon 399 of the X-ray repair cross-complementing group 1 (XRCC1) gene may subtly alter structure of DNA repair enzymes and modulate the repair capacity. Impaired DNA repair can lead to the development of cancers such as prostate cancer (PCA). Although the association between the XRCC1 codon 399 polymorphism and PCA risk. We performed a literature search of the PubMed database to identify all eligible articles that reported this association. Odds ratios (ORs) with 95% confidence intervals (CI) were evaluated to assess the association.We conducted an updated analysis of 18 case–control studies to determine the association between the XRCC1 codon 399 polymorphism were found . Moreover, subgroup analysis based on ethnicity revealed similar significant associations in Asians . Egger’s test did not reveal the presence of a publication bias.Significant associations between PCA risk and XRCC1 codon 399 polymorphism and PCA risk. Further carefully designed studies should be performed.Our updated analysis provides evidence for significant association between  Prostate cancer (PCA) is the most diagnosed cancer and has the second highest mortality in USA.8XRCC1. Among these, the codon 399 polymorphism has been wildly reported to be associated with PCA risk.14The X-ray repair cross-complementing group 1 (XRCC1) gene is located at 19q13.2 and encodes a multi-domain protein that acts as a scaffolding intermediate between ligase III, DNA polymerase-β, and poly-ADP-ribose polymerase.XRCC1codon 399 polymorphism on in the development of PCA. Here, we performed an updated meta-analysis to estimate the association between the XRCC1codon 399 polymorphism and PCA risk.To date, there have been 18 case–control studies in 15 articlesXRCC1 codon 399 polymorphism and PCA risk. Eligible studies were found by searching PubMed for relevant reports published between 2002 and 2012. The search terms were “XRCC1” or “X-ray repair cross-complementing group 1,” “polymorphism” or “variant,” and “prostate cancer” or “prostate.” A total of 32 articles were retrieved, of which 15 studies reported on the association between XRCC1 codon 399 polymorphism and PCA risk.We tried to include all case–control studies published to date about the association between XRCC1 codon 399 polymorphism and PCA risk; (2) case–control study; (3) available genotype frequency; (4) English language; and (5) full-text manuscript.(1) association between the (1) no control population; (2) no available genotype frequency; and (3) duplicated studies .Data included the following: first author, publication year, country, ethnicity, source of control, each genotype frequency of the case and control groups, genotype methods, and the Hardy–Weinberg equilibrium (HWE) value of the controls.XRCC1codon 399 polymorphism and PCA risk. The association between XRCC1codon 399 and PCA risk was determined by 3 different models: allelic contrast , homozygote comparison (AA vs. GG), and the recessive model (AA vs. AG+GG). Subgroup analysis was performed based on the ethnicity and source of case subgroups.Odds ratios (ORs) with 95% confidence intervals (CI) were used to measure the strength of the relationship between the Q-test, and the statistical significance of the summary OR was determined with the Z-test. Heterogeneity was ruled out when P > 0.05 for the Q-test; for such studies, the fixed effects model was used, and for other studies, the random effects model was used.P < 0.05 was considered significant. Sensitivity analysis was performed on excluded individual studies to assess the stability of the results. Publication bias was assessed by both Egger’s test and Begg’s test.Heterogeneity among the studies was evaluated with a chi-square-based Of the 32 abstracts retrieved in the PubMed search, 17 did not fulfill the criteria and were excluded. The 15 articles included in the study accounted for 18 case–control studies, which together comprised 4,479 cases and 4,281 controls . Details26P = 0.011 for heterogeneity; homozygote comparison: OR = 1.27, 95% CI = 1.04–1.56, P = 0.062 for heterogeneity; the recessive model: OR = 1.31, 95% CI = 1.10–1.57, P = 0.093 for heterogeneity)  . In the Sensitivity analysis was used to determine whether differences in the inclusion criteria of the different studies affect the results. No other single study influenced the summary OR qualitatively . Begg’s XRCC1 was the first human BER pathway gene to be cloned, and cells lacking this gene product are hypersensitive to ionizing radiation.XRCC1 that correlate with phenotypic changes have been identified.XRCC1 is A399G or R194W, located in the linker region separating the NH2-terminal domain (NTD) from the central BRCT1 (BRCA1 C terminus) domain. This linker region was also suggested to be a potential binding domain for several interacting proteins and is rich in basic amino acids. The substitution of arginine with hydrophobic tryptophan may affect the protein binding efficiency. The present meta-analysis examined 4,479 PCA patients and 4,281 healthy controls to evaluate the association between the XRCC1codon 399 polymorphism and PCA risk.DNA repair systems play an important role in protecting the genome from permanent damage by endogenous and exogenous mutagens, and impairment of these systems has been reported to increase the risk of various types of cancer, including PCA. At least four DNA repair pathways operate on specific types of damaged DNA: base excision repair (BER), nucleotide-excision repair (NER), mismatch repair (MMR), and double-strand break repair.XRCC1codon 399 polymorphism and PCA risk is affected by ethnicity. Significantly strong associations were found between the XRCC1codon 399 polymorphism and PCA in Asians but not in Mixed, Caucasians, or Africans. This suggests that this polymorphism occurs at different frequencies among various ethnic groups and could be considered a biomarker. This difference in distribution could explain the lack of well-replicated results across patient populations of different ethnicities.Our main finding is that the association between the XRCC1codon 399 polymorphism and PCA risk. Future studies should focus on these two ethnicities. Second, gene/gene, gene/environment interactions, and even interactions between different polymorphisms, should be included. The stage (TNM and Gleason score) and characteristic (PSA) of PCA should be included if possible. Finally, publication bias was detected, which may influence the power of results.Some limitations of our meta-analysis should be mentioned and addressed. First, there were only two Mixed or African case–control studies on XRCC1codon 399 polymorphism could increase PCA risk in Asians. To fully understand the influence of the XRCC1codon 399 polymorphism on susceptibility to PCA, as well as the role of genetic factors in the physiopathology of this disease, further studies in large, standardized, and ethnically diverse populations are needed.We provide evidence that


To evaluate the association between angiotensin I-converting enzyme insertion/deletion (ACE I/D) gene polymorphism and retinal vein occlusion (RVO). A total of 80 patients with retinal vein occlusion who was admitted to the Eye Department of Kartal Training and Research Hospital between 2008 and 2011, and 80 subjects were enrolled in this retrospective case–control study. Patients who experienced RVO within one week to six months of study enrolment were included, and those with coronary artery diseases, prior myocardial infarction history and coagulation disturbances were excluded from the study. The diagnosis was made by ophthalmoscopic fundus examination and fluorescein angiography. The ACE gene I/D polymorphism was determined by polymerase chain reaction, and the ACE gene was classified into three types: I/I, I/D and D/D. In multivariate logistic regression analysis, ACE D/D genotype (p = 0.035), diabetes-mellitus (p = 0.019) and hypertension (p = 0.001) were found to be independent predictive factors for RVO. The results of the present study reveal that ACE D/D polymorphism is an independent predictive factor for RVO. However, one cannot definitely conclude that ACE gene polymorphism is a risk factor for retinal vein occlusion. Angiotensin I-converting enzyme (ACE), dipeptidyl peptidase, is a membrane-bound enzyme, which is present in endothelial and epithelial cells of various tissues, and innards including lungs and kidneys. Angiotensin I-converting enzyme converts Angiotensin I to Angiotensin II, a very potent vasoconstrictor agent . AngiotePrevious studies have reported that plasma levels of angiotensin II are closely associated with ACE insertion/deletion (I/D) polymorphism and that the serum level of ACE is likely to increase two-fold in the presence of ACE D/D polymorphism, consequently increasing the levels of plasma angiotensin II . It has There are very few studies examining the relationship between ACE gene polymorphism and retinal vein thrombosis, with controversial results -14. TherThis case–control multi-center study composed of 80 patients, who experienced RVO one week to six months before enrolment. Control group composed of age and sex matched 80 persons in which retinal vein occlusion, other ocular diseases excluded with detailed ocular examination who referred to the eye clinic from internal medicine clinics in which study conducted. Patients gave written informed consent in accordance with the Declaration of Helsinki. The Institutional Review Board of Kartal Koşuyolu Heart Training and Research Hospital approved the study.The patients with RVO and controls underwent a general physical examination, and a thorough cardiovascular and ophthalmic examination. A detailed medical history was taken from the study cohort. We excluded the subjects who had diabetic and/or hypertensive retinopathy findings among the controls. This is because subjects having vascular changing related to diabetes mellitus (DM) and/or hypertension (HT) might cause confusion while being evaluated the retinopathy related to RVO.The diagnosis of RVO was made by ophthalmoscopic fundus examination and flourescein angiography. On the fundus examination, disc swelling, venous dilation or tortuosity, retinal hemorrhages, cotton wool spots and on the flourescein angiography demonstrating extensive areas of capillary closure, venous filling defects and increased venous transit time were assesed as the diagnosis of RVO by the same ophthalmologist.The patients and controls were assessed for coagulation abnormalities and thrombosis .Patients with abnormalities in coagulation parameters, previous thrombosis and family history of thrombosis, using oral contraceptives and hormone replacement therapy, having renal and coronary artery diseases and a prior history of myocardial infarction were excluded.Hypercholesterolemia was defined as a total serum cholesterol level >200 mg/dL on admission or maintenance of normal cholesterol levels with statin therapy. Hypertension was defined as a systolic blood pressure ≥140 mmHg and a diastolic blood pressure ≥90 mmHg or current use of antihypertensive medications. Patients on oral anti-diabetic drug (OAD) therapy and/or insulin were considered as having diabetes.Polymerase chain reaction (PCR)–cDNA coagulation measures were performed to detect ACE polymorphisms in cases with RVO and in controls. Blood samples were taken from the antecubital vein after an overnight fasting. Whole blood samples from the patients were collected in ethylenediaminetetraacetic acid (EDTA) tubes. Total genomic DNA was isolated from whole blood samples using GenXtract DNA extraction system according to the manufacturer’s instructions (Vienna Lab Diagnostic GmbH). Then, target DNA regions were amplified by multiplex polymerase chain reaction (PCR) using biotinylated primers. Thereafter, amplified products were separated on 3% agarose gel. After observing the amplicons of the related genes, the amplified products were hybridized to a test strip containing allele specific nucleotide probes immobilized on a nitrocellulose membrane, using Cardiovascular Disease (CVD) Strip Assay (Vienna Lab Diagnostic GmbH). Hybridization process was performed with Tecan Profiblot T48 hybridization device. Bound biotinylated sequences were detected using streptavidin-alkaline phosphatase and color substrates.Angiotensin I-converting enzyme gene was classified as I/I, I/D, D/D. The ACE gene I/D polymorphism was determined by PCR, using a primer pair flanking the polymorphic region of intron 16 that produces an amplified 490-bp , a 190-bp product  or both. All the reactions were performed according to the method by described previously . The allContinuous variables were expressed as mean ± standard deviation. Categorical variables were expressed as percentages. The group means of continuous variables were compared using an independent samples t-test. Categorical variables were compared using the chi-square or the Fisher’s exact tests. Multivariate logistic regression analysis was applied to identify the independent predictors of RVO. Variables which identified as significant in the univariate analysis  were included in the model. Two-tailed p values <0.05 were considered to indicate statistical significance. The Statistical Package for the Social Sciences  version 11.5 was used for all statistical analyses.This case–control study included 80 RVO patients  with a mean age of 60.2 ± 12.1 years, and 80 controls  with a mean age of 59 ± 12.6 years. Baseline demographic characteristics of the patients and control subjects are shown in Table Angiotensin I-converting enzyme D/D gene polymorphism was significantly higher in patients with RVO , while ACE I/D polymorphism was significantly lower in patients with RVO than that of the controls . However, with regard to ACE I/I gene polymorphism, there was no significant difference between the two groups , in contrast ACE I/D polymorphism was significantly higher in hypertensive controls than patients with RVO . With regard to ACE I/I gene polymorphism, there was no significant difference between the two groups .Frequency of ACE DD polymorphism in diabetics of control group was significantly lower compared to that of patients with RVO . ACE I/D polymorphism was significantly higher in the diabetic controls than patients with RVO . However, with regard to ACE I/I gene polymorphism, there was no significant difference between the two groups .Angiotensin I-converting enzyme D/D genotype [Odds Ratio (OR) = 3.83, 95% Confidence Interval (CI) = 1.10-13.37, p = 0.035, diabetes mellitus  and hypertension  were found to be the independent predictive factors for RVO in multivariate logistic regression analysis.Current smoking, hyperlipidemia and ACE I/D genotype were the significant variables in the univariate analysis; however, these parameters lost significance in multivariate analysis Table .The main finding of the present study is that ACE D/D gene polymorphism is an independent predictive factor for RVO. ACE I/D gene polymorphism was found to be significantly lower in patients with RVO, when compared with controls; however, ACE I/I polymorphism was similar between the two groups.Retinal vein occlusion is the second most common retinal vascular disease after diabetic retinopathy in the elderly. It leads to serious visual loss and blindness. RVO is classified as central (CRVO) and branch RVO (BRVO) depending on the site of occlusion. The prevalence of BRVO (0.6 - 1.6%) is greater than that of CRVO (0.1 - 0.4%) ,14. The The potential risk factors related to RVO include age, smoking, diabetes, hypertension, hyperlipidemia and open-angle glaucoma -22. In aAngiotensin II promotes vasoconstriction, inflammation and thrombosis, and as a proatherogenic factor, decreases fibrinolytic activity via increasing PAI-1 levels. Previous studies have shown that the plasma level of angiotensin II is closely associated with ACE gene polymorphisms. ACE I/D gene polymorphism has been described as an independent risk factor for thrombotic diseases as well -10.On the other hand, many earlier studies have associated ACE I/D gene polymorphism with diabetes mellitus and hypertension, and stated that the results are specific to the community studied, and ethnicity -27. TherIn our study, the ACE D/D gene polymorphisms, diabetes mellitus, hypertension, hyperlipidemia, and smoking were found to be significantly higher in the RVO group compared to controls. Gori et al.  investigAs a result, in this study, the ACE D/D gene polymorphism was found to be an independent predictive factor for RVO. However, it is difficult to say that this polymorphism is a definite risk factor for RVO, due to the scarcity of large-scale studies evaluating the distribution of ACE I/D gene polymorphism in patients with diabetes mellitus and/or hypertension among Turkish population. Therefore, studies performed on the relationship between RVO and ACE gene polymorphism should take into consideration other systemic risk factors for RVO, keeping in mind that the results obtained would be specific to the population studied. This will yield more meaningful results that will help develop new prophylactic and therapeutic strategies for the diagnosis, treatment and follow-up of RVO.The authors declare that they have no competing interests.IK drafted the manuscript, involved in conception, design, acquisition, analysis and interpretation of data. AK participated in the design of the study and performed the statistical analysis. TB conceived of the study, and participated in its design and coordination and helped to draft the manuscript. VO performed the statistical analysis, drafted the manuscript. AE involved in conception and design, acquisition of data.CD analysis and interpretation of data.AE analysis and interpretation of data.CK involved in conception and design, acquisition of data.YÖ participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.


The aim of this study was to identify environmental or genetic risk factors that are associated with invasive meningococcal disease (IMD) in children in Greece.MBL2 gene and quantitative measurement of MBL serum protein levels were performed using Nanogen NanoChipR 400 technology and immuno-enzyme techniques, respectively.A case-control study was performed in 133 children (44 cases and 89 controls) aged between 0–14 years, who were hospitalized in a children's hospital in Athens. Demographics and possible risk factors were collected by the use of a structured questionnaire. To investigate the association of mannose binding lectin (MBL) with IMD, a frequency analysis of the haplotypes of the 2 (OR = 3.16), were independent risk factors associated with IMD. Overall 18.8% of patients had a MBL2 genotype with low functionality compared to 10.1% of healthy controls, but this was not statistically significant (p = 0.189).The multivariate analysis revealed that changes in a child's life setting , paternal smoking (OR = 4.51), upper respiratory tract infection within the previous month (OR = 3.04) and the density of people in the house/100mPrevention strategies aimed at reducing parental smoking and other risk factors identified in this study could decrease the risk of IMD among children in Greece. Neisseria meningitidis), a Gram-negative bacterium that is classified into 13 capsular groups according to its capsular polysaccharides. Six of these , are of clinical significance as they cause invasive infections. In Europe, groups B and C are mainly responsible for IMD [Invasive meningococcal disease (IMD) is a contagious bacterial disease caused by a meningococcus  forms [Meningococcal infections are transmitted between people through respiratory droplets or secretions.  no harm –7. There no harm –10. Seveb) forms . In GreeThe risk of meningococcal infection in an individual is dependent on the balance of the virulence of the strain and the host’s immune response. Moreover, several environmental risk factors have been associated with the disease in several countries –13. BothMBL2 gene.Genetic mutations of MBL(an acute protein phase that contributes to the elimination of bacteria by activating the complement system) have been identified as possible risk factors associated with IMD in several studies –25. As cApproval of the study protocol was received by the Ethics Committee of Aghia Sofia Children’s Hospital, which waived the need for written consent. Parents or guardians were informed about the aim of the study, and they provided written consent for their child’s participation in the study.A case-control study was performed using 133 children (44 cases and 89 controls). All participants were children aged between 0–14 years, who had been hospitalized in two children's hospitals (Aghia Sophia and P & A Kyriakou) in Athens, Greece, within a 2-year period from January 2011 to December 2012.N. meningitidis was identified in samples of biological material (blood or CSF) in the laboratory and isolated using bacterial cultures or molecular techniques, such as polymerase chain reaction (PCR). In addition, to increase the reliability and the power of our genetic research, the frequency analysis of the gene polymorphisms of MBL2, included 45 extra blood samples from the National Reference Centre for Meningitis (NRCM). These samples were collected from cases of IMD in children aged 0–14 years, who had become ill during the same period (2011–2012) and had been admitted to various hospitals across Greece. Our study was designed to encompass two spring seasons in order to capture any respective seasonal increase in IMD.Cases were had been hospitalized with a diagnosis of IMD . In all cases, Controls were children hospitalized in the surgical wards of the two hospitals with a diagnosis that was unrelated to IMD or other infections. All controls were matched to cases using the sex and age (year of birth) of each child, and the week of admission. When it was possible, at least two controls were randomly selected for every case.MBL2 haplotypes and the serum levels of MBL protein. For each blood sample, 0.5 mL was stored immediately at -80°C, in order to be used in the frequency analysis of MBL2 polymorphisms. After centrifugation of the remaining blood sample, plasma was collected and kept readily cryopreserved at -80°C for the quantitative measurement of MBL.In both cases and controls, a whole blood sample (3–4 mL) was collected on the day the child was admitted to the hospital, in order to study the A questionnaire was distributed to the parents of both cases and controls, in order to obtain information on the following: their child’s demographic details ; family history ; housing environment . Moreover, the questionnaire included questions regarding perinatal history and breastfeeding; history of hospitalization; vaccination history; medications/ social history during the last month: attendance at nursery school, elementary/high school/college attendance; participation in sporting events; attendance at parties and playgrounds; church attendance; visits to restaurants or coffee bars; kissing other people; use of public transport; and change of life setting (relocation or vacation). If the answer to the question of change in life setting was positive the parents were asked to specify. Parents were also asked to report if their children had signs and symptoms compatible with upper respiratory infection (fever and/or cough and/or sore throat and/or rhinitis) during the last month. It should be noted that some questions in the questionnaire  were relevant to older children (≥12 years old), while other questions  were relevant to children aged ≤10 years.MBL2 includes three polymorphic sites in exon 1  forming the haplotypes AO, OO, and AA, three polymorphisms in the area of the promoter  forming the haplotypes HY, LY and LX and four polymorphic sites located in exon 4 (carbohydrate recognition domain-CRD). The frequency analysis of MBL2 polymorphisms was performed by the Choremeio Research Laboratory of Medical Genetics at the University of Athens. Of the 44 cases of IMD, only 36 agreed to participate in the genetic analysis while 69 out of the 89 of the controls agreed to participate . To increase the power of the study, 45 cases of IMD from NRCM were included in the genetic analysis to achieve a total number of 81 cases of IMD analyzed.MBL2, we developed an advanced high throughout methodology using the Nanogen NanoChipR 400 system . Three separate regions of the MBL2 gene containing SNPs were designed for PCR amplification using the PrimerQuestSM tool provided by IDTR (Integrated DNA Technologies). The set-up of all PCR reactions was performed automatically using the Biorobot 3000 platform  and carried out in a Techne TC-412 thermal cycler using the amplicon-down format of the Nanogen protocol. Results were estimated by the instrument's software.Genomic DNA was isolated from 350 μl of peripheral blood, using the BioRobotR M48 System  and the MagAttractR DNA Blood Midi M48 Kit . For the characterization of the six SNPs in The quantitative measurement of the MBL levels of serum was performed in 43 of 44 (97.7%) patients, and in 73 of 89 (82.0%) controls at the Department of Hygiene and Epidemiology at the University of Thessaly, using the MBL Oligomer ELISA Kit (BioPorto Diagnostics Co.).All data collected from the study participants  were entered into an electronic database using Epi-info software . Statistical analysis was performed using IBM SPSS Statistics software .2), which was used to distinguish between IMD cases and controls.Quantitative variables were presented either as mean values with standard deviation or as a median value with the interquartile range (IQR). Qualitative variables were presented as absolute and relative frequencies with the corresponding 95% confidence intervals (95% CI). The receiver operating characteristic (ROC) curve analysis was conducted to determine the optimal cutoff values of the quantitative variable density  and IMD. The Mann-Whitney test was used to explore differences between IMD cases and controls with regards to quantitative non-normally distributed variables.In the multivariate analysis, multiple logistic regression analysis was performed, using the backward stepwise conditional method with a removal criterion of p-value equal to or greater than 0.10, inorder to identify the independent risk factors for the onset of IMD by calculating the odds ratios (ORs) and the corresponding 95% CI. The dependent variable was IMD and as independent variables were used all the statistically significant risk factors found in the univariate analysis together with age and gender were used as independent variables. For the genetic analysis of MBL, the necessary sample size was calculated to be 200 children (100 per group) assuming a study power of 0.80, alpha 0.05, and considering 10.0% of low MBL in controls to identify a significant OR 3.00 (cases vs. controls). A result with a p-value <0.05 was considered to be statistically significant.During the initial study period, 50 cases of IMD were hospitalized in both hospitals. In two cases, the diagnosis was not confirmed and the guardians of four patients  refused to participate in the study. Overall, 44 cases of IMD participated in the study (response rate: 91.1%), of whom 27 (61.3%) were males and 17 (38.7%) females. The median age was 3 years (IQR: 1–4 years). Regarding race, 93.2% of them were white, 2.3% black, and 4.5% were from an other racial background. Also, 18 IMD cases (~41%) were second-generation immigrants, of whom 13 (~72%) were born in Greece while their parents were from Albania. Seven patients presented with septicemia, nine had meningitis and 28 patients had both. Sixteen patients were diagnosed using PCR and 28 through culture tests. The majority of cases were caused by group B meningococcus (36/44), one patient had group A and one had group W, while meningococcus was nongroupable in six patients. Furthermore, 23 out of 40 (57.5%) cases of IMD had been vaccinated with meningococcal meningitis C vaccine.Regarding the controls, 99 patients were initially recorded but only 89 controls participated in the study (response rate: 89.9%). Out of these 89 controls, 65 (73%) were males and 24 (27%) were females, while 98.8%were white, 0% were black and 1.2% had another racial background. The median age of controls was 3 years (IQR: 1–5 years). Approximately 24% of the controls were second-generation immigrants.N. meningitidis type C, or parental occupation, were found to have any statistically significant association with the occurrence of IMD. Moreover, factors such as the use of antibiotics, participation in sports activities, participation in parties, or kissing other people during the previous month were not statistically significant. Finally, no statistically significant difference was found in the serum protein levels of MBL between cases and controls.All statistically significant risk factors, according to the univariate analysis, are presented in As shown in Parental smoking  and paternal smoking  were identified to be associated with IMD, although the difference for maternal smoking was not statistically significant . Moreover, for fathers a dose-response was revealed between number of cigarettes per day and therisk of IDM. The OR for smoking ≥20 cigarettes per day was 3.32 (95% C.I.: 1.46–7.58), compared to 2.56 (95% C.I.: 0.62–10.53) for smoking 1–19 cigarettes per day. The mean number of cigarettes per day smoked by the fathers was double of that of the mothers (20 vs. 10).As shown in MBL2 gene, except for one sample. The low functionality of MBL2, was observed in 18.8% of the patients and in 10.1% of the control group, but this difference was not statistically significant (p = 0.189) , moderate  and low (LXA / O and O / O) functionality of the = 0.189) .N. meningitidis. It is known that the risk of invasive IMD is higher in newly colonized people [This study revealed that a recent history of relocation or holiday, paternal smoking, a recent history of viral upper respiratory tract infection and crowded home conditions were independent risk factors for IMD. One interesting and possibly important novel finding of our study was the association between a change in the life setting either by relocation or vacation in the previous month with IMD (OR = 7.16). During these trips, extensive social activities are expected together with changes in pharyngeal flora as indicated by previous studies . The pred people . This fid people .N. meningitidis has also been documented in previous studies. It has been suggested that this increased risk may be attributed to the increased ability of the bacteria to adhere to mucosa in the presence of smoke [Our findings are in line with well-known risk factors for IMD, such as a history of viral respiratory tract infections , and parof smoke .Finally, multiple logistic regression analysis has revealed that the crowd density in the house (number of persons per 100 square meters of the house) was an independent risk factor for IMD. It should be noted that a nationwide population-based case-control study among preschool children reported that the risk of IMD increased with an increasing household density . A numbeMBL2 gene and its predisposition to IMD. The patients were classified into three groups of high, moderate and low functionality of the MBL2 gene, based on the combination of polymorphisms in exon 1 and in the region of the promoter. Patients mostly had the low functionality genotype, compared to healthy controls  several exposures more readily than controls. However, on the other hand, it is not likely that parents of cases over-reported their smoking habits, and consequently, the exposure of their children to passive smoking. On the contrary, it would be expected that parents would under-report their smoking activities, which could lead to an underestimation of the impact of parental smoking on the risk associated with IMD. A further limitation of our study was that symptoms that were suggestive of an infection of the upper respiratory tract were based on self-reports, without the implementation of serological tests for the detection of viral antigens. Finally, we matched cases to controls by age and thus we lost the opportunity to verify the age as a risk factor. However, as age is a well-known strong confounder, we preferred to control for age to be able to study the other possible risk factors.In conclusion, our case-control study indicated that paternal smoking, a recent history of upper respiratory tract viral infection, crowded households and recent relocation/vacation activities were independent risk factors for IMD. Additional studies are needed to explore in detail the role of relocation or holidays as risk factors for IMD and to assess the actual risk posed. Preventive activities aimed at reducing parental smoking and other risk factors could decrease the risk of IMD among children in Greece.


The Rho GTPase-activating protein 35 (ARHGAP35), an important Rho family GTPase-activating protein, may be associated with tumorigenesis of some tumors. Here, we investigated the relationship between an important polymorphic variant at 3′-UTR of this gene (rs1052667) and osteosarcoma risk and prognosis. This hospital-based case-control study, including 247 osteosarcoma patients and 428 age-, sex-, and race-matched healthy controls, was conducted in Guangxi population. Genotypes were tested using TaqMan PCR technique. We found a significant difference in the frequency of rs1052667 genotypes between cases and controls. Compared with the homozygote of rs1052667 C alleles (rs1052667-CC), the genotypes with rs1052667 T alleles  increased osteosarcoma risk . Moreover, rs1052667 polymorphism was correlated with such pathological features of osteosarcoma as tumor size, tumor grade, and tumor metastasis. Additionally, this polymorphism also modified the overall survival and recurrence-free survival of osteosarcoma cases. Like tumor grade, ARHGAP35 rs1052667 polymorphism was an independent prognostic factor influencing the survival of osteosarcoma. These results suggest that ARHGAP35 rs1052667 polymorphism may be associated with osteosarcoma risk and prognosis. Osteosarcoma is the most frequent primary malignant bone tumor and usually occurs in patients between 10 and 25 years of age . HoweverTherefore, a better understanding of its basic biology is urgently needed to identify its risk and prognostic markers. Several studies have reported potential associations of common genetic variants with osteosarcoma risk in biologically plausible pathways. This suggests that the genetic factors could play important roles in the pathogenesis of this malignant tumor , 6.The Rho GTPase-activating protein 35  is an important Rho family GTPase-activating protein, and is identified as a tyrosine-phosphorylated protein associated with p120RasGAP in v-Src transformed cells –14. FuncThe present study was approved by the ethics committees of the hospitals involved in this study. This hospital-based case-control study was conducted in Guangxi Zhuang Autonomous Region, China, a relatively high incident area of osteosarcoma. All osteosarcoma patients and control individuals were residents of Guangxi Zhuang Autonomous Region and were recruited from the affiliated hospitals of Guangxi Medical University between January 1996 and August 2005. All cases were histopathologically confirmed. During the same period, control subjects without a history of cancer were randomly selected from a pool of healthy volunteers who visited the general health check-up center of the same hospitals because of their routine scheduled physical exams , 23. To Among 247 osteosarcoma cases, about 57 percent (140/247) of osteosarcomas located in femur. All cases received surgical resection of primary tumor; however, only fifty-seven cases underwent the radical treatment (including both curative resection and adjuvant chemotherapy). In this study, tumor grade was evaluated according to Broders' grading system .Leukocytes were isolated from peripheral venous blood samples from all tumor patients and control subjects by standard procedures. DNA was then extracted from leukocyte samples by standard phenol-chloroform extraction and ethanol precipitation. DNA samples were stored at −20°C until additional analysis.µL consisting of 1 × TaqMAN Universal Master Mix II , 1 × TaqMan SNP Genotyping Assay Mix , and about 75 ng of genomic DNA. Cycling conditions were 95°C for 30 s, and 50 cycles of 95°C for 15 s, and 60°C for 1 min. For quality control, laboratory personnel were blinded to case and control status. Controls were included in each run, and repeated genotyping and sequencing of a random 20% subset yielded 100% identical genotypes.The ARHGAP35 rs1052667 genotypes were analyzed by TaqMan polymerase chain reaction (PCR) on an iCycler iQ real-time PCR detection system . The corresponding TaqMan SNP Genotyping Assay Kit (cat# 4351379) was obtained from Applied Biosystems, Carlsbad, CA, USA. TaqMan PCR was performed in total volume of 25 For survival analysis, we followed all osteosarcoma cases. All patients underwent serial monitoring every 2 months for the first 2 years and semiannually thereafter for detection of any recurrence. In this study, the last follow-up day was December 31, 2013, and survival status was confirmed by clinic records and either patient or family contact. The duration of overall survival (OS) was defined as from the date of curative treatment to the date of death or last known date alive, whereas the recurrence-free survival (RFS) was defined as from the date of curative treatment to the date of tumor recurrence or last known date alive.χ2 test or Fisher's exact test was used to test the differences between osteosarcoma patients and control subjects in the distribution of gender, age, race, and ARHGAP35 rs1052667 genotypes. Because this study was based on an individually matched design, conditional logistic regression was used to evaluate odds ratios (ORs) and 95% confidence intervals (CIs) for risk of osteosarcoma. Kaplan-Meier survival analysis (with the log-rank test) was used to elucidate the relationship between ARHGAP35 rs1052667 polymorphism and osteosarcoma prognosis. Hazard ratios (HRs) and 95% CIs for ARHGAP35 genotypes were calculated from a multivariate Cox regression model (with stepwise forward selection based on the likelihood ratio test). In the present study, a P  value of < 0.05 was considered statistically significant.All analyses were performed with the statistical package for social science (SPSS) version 18 . Pearson's P > 0.05).In this study, 247 osteosarcoma cases and 428 controls were included in the final analysis. The demographic characteristics of all cases and controls are shown in P < 0.01), resulting in an Ser allele frequency of 32.0% in cases and 11.9% in controls. Logistic regression analysis exhibited that the adjusted OR for osteosarcoma for these individuals carrying rs1052667-CT compared with those exhibiting the homozygote for C alleles (rs1052667-CC) was 2.41  and the corresponding OR for those featuring rs1052667-TT was 7.35 . These results showed that osteosarcoma risk was associated with the number of rs1052667 T alleles.Pinteraction > 0.05; To evaluate possible interactive effects of matching factors  and ARHGAP35 rs1052667 polymorphism on osteosarcoma risk, we performed a series of bivariate stratified analyses by matching factors . BecauseP is 1.19 × 10−11 for OS and P is 2.04 × 10−17 for RFS, resp.; Figures To investigate the effects of ARHGAP35 polymorphism on outcome of osteosarcoma patients, we followed all cases and analyzed the survival information of all osteosarcoma cases. During the follow-up period of these patients, 222 faced tumor recurrence with 15.9% of the 5-year RFS rate, and 238 died with 12.5% of the five-year OS rate. Kaplan-Meier survival analysis showed that patients with ARHGAP35 rs1052667 T alleles featured a significantly poorer prognosis than those with rs1052667-CC  or the nonrisk genotype (rs1052667-CC) and the clinical pathological characteristics of osteosarcoma was performed separately. Results showed that these osteosarcoma cases with risk genotypes of ARHGAP35, compared to those without risk genotypes, faced larger tumor size (OR is 4.85), lower tumor differentiation (OR is 4.07), and higher metastasis risk . These results imply that this polymorphism may have functional significance in osteosarcoma carcinogenesis.Osteosarcoma is one of major cancer types in the Guangxi Zhuang Autonomous Region; the possible risk factors of which include radiation exposure, foreign bodies, genetic predisposition, and so on. Increasing epidemiological evidence has shown that an individual susceptibility related to genetic factors might be associated with osteosarcoma carcinogenesis , 6.2-terminal GTP-binding domain (GBD), (2) a middle domain (MD), and (3) a COOH-terminal GAP domain, which displays specificity for GTP-bound RhoA [While ARHGAP35 spans 87 kb on chromosome 19q13.3 and contains 7 exons and 6 introns (PubMed Databases). Its encoding protein is a 190 kDa protein consisting of three major functional domains: (1) an NHund RhoA . Functiound RhoA , 27. Recund RhoA , 28. In http://www.ncbi.nlm.nih.gov/SNP/snp_ref.cgi?rs=rs1052667). However, higher frequency was observed in the individuals with osteosarcoma, and following analysis showed this polymorphism increased osteosarcoma risk. These results suggested ARHGAP35 rs1052667 polymorphism might modify the risk of tumors such as osteosarcoma.With the Human Genome Project developing, more than one hundred polymorphisms have been identified in ARHGAP35 (dbSNP in NCBI Database). In this study, we only analyzed ARHGAP35 rs1052667 polymorphism, primarily because this polymorphism is relatively common in most populations, whereas other polymorphisms are rare. In this study, we collected 247 osteosarcoma and 428 control samples from Guangxi Zhuang Autonomous Region, a relatively high incident area of osteosarcoma. we found that about 20 percent of control individuals had ARHGAP35 rs1052667 T alleles, similar to the data from the Human Genome Project  may have implications for screening and prevention.


MTHFR C677T polymorphism has been indicated to be a risk factor for cancers, but its association with head and neck cancer (HNC) risk remains inconclusive. In the present study, we aimed to get a more precise estimation by performing a quantitative meta-analysis. Published papers up to Jun 2014 was searched and screened. Necessary information was rigorously extracted for data pooling and analyzing, and then, subgroup analyses on ethnicity, source of controls, sample size, tumor type, smoking and drinking status were also carried out. As a result, twenty-three case-control studies including 14298 subjects were included. The overall data failed to reveal a significant association between MTHFR C677T polymorphism and HNC risk . However, in the subgroup analysis about drinking status, increase risk was shown in the heavy drinking subgroup . In conclusion, the results of the present study suggest that Homozygous TT alleles of MTHFR C677T polymorphism might be a risk factor for HNC among individuals who have a heavy drinking history. Further studies are needed to get a more definitive conclusion. Head and neck carcinoma (HNC), the sixth most frequent kind of cancer worldwide, is a group of biologically similar cancers that originate from head and neck regions such as oral cavity, pharyngeal cavity, and larynx2de novo deoxynucleoside synthesis and for intracellular methylation reactions in humansMTHFR), a key enzyme for intracellular folate hemeostasis and metabolism, catalyses the irreversible conversion of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate that is the primary circulating form of folate, providing methyl groups for the methylation of homocysteine to methionine6MTHFR (rs1801133), C677T in exon 4 , may be implicated in tumorigenesis with alteration of MTHFR enzyme activityMTHFR enzyme activity of the 677CC wild-type genotypeGenerally, folate plays a fundamental role of providing methyl groups for MTHFR rs101133, C6MTHFR polymorphism with HNC have generated inconclusive results. Clarifying this association may help us better understand the possible risk of HNC and therefore contribute to its prevention. Previously, we assessed the relationship between MTHFR polymorphism and oral cancer riskPublished data on the association of methylenetetrahydrofolate reductase, MTHFR, head and neck, oral, pharynx, larynx, thyroid, mouth, neoplasm, tumor, cancer, variation and polymorphism. All searched studies were retrieved and the bibliographies were checked for other relevant publications. Review articles and bibliographies of other relevant studies identified were hand searched to find additional eligible studies.The meta-analysis was presented according the PRISMA-MOOSE statement. We carried out a search in the internet covering well-known biomedical databases such as Medline, EMBASE, and CNKI without a language limitation. Papers published up to Jun 1, 2014 were included. The literature selection was performed from Jun 1, 2014 to Jun 30, 2014. The following keywords were used for searching: MTHFR C677T polymorphism with HNC risk; second, studies should be case-control or cohort designed; third, papers must offer the size of the sample, odds ratios (ORs) and their 95% confidence intervals (CIs), the genetic distribution or the information that can help infer the results. After deliberate searching, we reviewed all papers in accordance with the criteria defined above for further analysis.The following criteria were used for the literature selection: First, studies should concern the association of Information was carefully extracted from all eligible papers by two of the authors (XZ and JS) independently according to the inclusion criteria mentioned above. If their results were conflicting, an agreement was reached following a discussion. If a consensus could not be reached, another author (DL) was consulted to resolve the dispute and then a final decision was made by the majority of the votes. The extracted data were entered into a database.MTHFR C677T polymorphisms and HNC risk was estimated for each study. The pooled ORs were performed for a homozygote comparison model (TT versus CC), a dominant model (TT + CT versus CC) and a recessive model (TT versus CT + CC), respectively. For detection of any possible sample size biases, the OR and its 95% confidence interval (CI) to each study was plotted against the number of participants respectively. For assessment of the heterogeneity between studies, two indexes were calculated. One was I2 metric, with I2 = 0-25% indicating no heterogeneity, I2 = 25-50% indicating moderate heterogeneity, and I2 > 50% indicating large heterogeneityP > 0.1 (indicating no heterogeneity), ORs were pooled according to the fixed-effect model (Mantel-Haenszel), Otherwise, the random-effect model (DerSimonian and laird) was used. The significance of the pooled ORs was determined by Z-test. The Hardy-Weinberg equilibrium (HWE) was assessed via Fisher’s exact test. Publication bias was assessed by visual inspection of funnel plotsThe odds ratio (OR) of Publications relevant to the key words were retrieved and screened originally. A total of ninety-seven studies were searched and screened for retrieval, of which sixty-seven irrelevant studies were excluded. Then, three review articles1516181922232425262728293031323334353637383940414243All the included studies were written in English except for one study in ChineseMTHFR genotype of the included studies were also presented , dominant model (TT + CT versus CC) and recessive model (TT versus CT + CC), respectively. As shown in MTHFR C677T polymorphism with HNC risk , indicating that TT carriers who have a heavy drinking history might have an increased HNC risk.To test the stability of the overall results, we first excluded the studies whose genetic distributions in controls markedly deviated from HWE, given that the deviation might result in any biasFunnel plots were created for assessment of possible publication bias. Then, Egger’s linear regression tests were used to assess the symmetry of the plots. As a result, the data suggest that the funnel plots were symmetrical for the three models , suggesting that the publication bias may have little influence on the results .MTHFR C677T polymorphism on cancer risk were controversial. Recent published meta-analyses showed that 677 C > T variation might contribute to the development of breast and esophageal cancer47MTHFR C677T polymorphism may play various roles in different carcinomas. In the present study, though the overall results failed to suggest a relationship between MTHFR C677T variation and HNC risk, the subgroup analysis indicated that homozygous 677TT might be associated with increased susceptibility to HNC in individuals who have a heavy drinking history.The effects of MTHFR 677T allele with HNC risk was observed among the three subgroups. The results were in line with the overall data, indicating that the effects of discrepancies among different ethnicities on the results were not evident.Considering the possible effects of the confounding factors on the overall data, we tried to carry out subgroup analyses. In subgroup analysis according to ethnicity, no significant association between When the data were stratified by source of controls and sample size, respectively, the results were not significantly different from the whole results, indicating that these two factors have little influence on the results. Likewise, in the subgroup stratified by tumor type, the results were in accordance with the overall data, implying that the dissimilarity of tumor types rarely affect the pooled results.MTHFR variation for HNC risk. However, when the data were divided by drinking status, elevated risk was shown in the heavy drinking subgroup under the homozygote comparison model. The data were in line with our previous meta-analysis about oral cancerMTHFR 677C → T polymorphism might play different roles for cancer risk according to the folate levels. It might reduce cancer risk when folate status is normal5558Whether chronic smoking interacts with folate status in the pathophysiologic process of disorders remains controversialEvident heterogeneity was shown in every model for the overall data, and thus, random-effect models were used to pool the data. Nevertheless, significant heterogeneities were removed in some subgroups while observed in other subgroups, indicating that the heterogeneities may be multifactorial. Besides the confounding factors considered in the subgroup analysis, other factors such as age, gender and life-style factors might contribute to the heterogeneities.Several limitations might be involved in this meta-analysis. First, only publications written in English and Chinese were searched and selected. It is possible that articles written in other languages that might meet the inclusion criteria were missed. Thus, inevitable publication biases might exist though the funnel plots were tested to be symmetrical. Second, subgroup analyses regarding age, gender and other risk factors such as virus infection status and betel quid chewing have not been conducted because the data in the primary literature were insufficient. Third, hospital-based controls were used in some of the included studies, leading to non-differential misclassification bias. Moreover, the controls in some studies were not strictly matched to the cases. Therefore, any selection bias might have an influence on the overall results and future investigations with large sample sizes are required.MTHFR C677T polymorphism with HNC risk, the subgroup analysis indicated that MTHFR 677TT alleles might increase HNC risk in individuals who have a heavy drinking history.Despite the limitations, though the overall data of the present meta-analysis did not reveal an association of How to cite this article: Zhuo, X. et al.MTHFR C677T polymorphism interaction with heavy alcohol consumption increases head and neck carcinoma risk. Sci. Rep.5, 10671; doi: 10.1038/srep10671 (2015).