Glaucoma is the leading cause of irreversible blindness in the world. Recent evidence indicates a role for genetic susceptibility to primary open-angle glaucoma (POAG). The relation between myocilin polymorphisms and POAG susceptibility has been studied in different populations.A meta-analysis of 32 published genetic association case-control studies, which examined the relation between POAG and the R46X, R76K, Y347Y, T353I, and Q368X polymorphisms of the myocilin gene, was carried out.In meta-analysis, significant associations were observed between POAG risk and two myocilin polymorphisms with summarized odds ratio of 4.68  for Q368X and 2.17  for T353I. Both Q368X and T353I were significantly associated with high-tension glaucoma, with summarized odds ratio of 4.26  and 2.26 (1.37–3.72). In Westerners, significant association was observed for Q368X mutation . However, in Asians it was for T353I .There is strong evidence that myocilin polymorphisms are associated with POAG susceptibility, and the prevalence of myocilin mutations might be ethnicity-dependent in Caucasians for Q368X and in Asians for T353I. Glaucoma, which causes optic nerve damage and visual field loss, is the leading causes of irreversible blindness worldwide The MYOC gene, also known as trabecular meshwork-inducible glucocorticoid response (TIGR) gene, was the first discovered to be linked to POAG in 1997 A total of 665 articles were identified across PubMed and Embase, and 57 full-text articles were retrieved. Finally, 32 studies met criteria and were included in the present meta-analysis 2 = 0.00%) was observed. There was no publication bias . In subgroup analysis by ethnicity, the association was significant in Caucasians, but not in Asians and Africans  , and no Africans . The ass2 = 0.00%). No publication bias was observed . Significant relation was also observed in Asians , 0.86 (0.69–1.08), and 1.02 (0.61–1.70) . There wDrosophila ocular hypertension model identified transcripts with altered regulation and showed induction of the unfolded protein response upon overexpression of transgenic human glaucoma-associated myocilin Myocilin is an eye protein found in the trabecular extracellular matrix, within the anatomic region that controls fluid flow, therefore, it might be the source of the resistance to aqueous humor outflow that causes the elevation of IOP in POAG Strength of the associations in the subgroup analyses with respect to study population was not consistent. In Caucasians, significant association was observed for Q368X mutation, and, in Asians, it was for T353I. Because the rate of Q368X mutation was rare, of being 0.6% in Asians and 0.3% in Africans respectively Normal tension glaucoma is a common form of open-angle glaucoma throughout the world, and yet there are many unanswered questions regarding both the mechanisms of the optic neuropathy. A certain level of IOP is the predominant causative risk factor in POAG, while additional IOP-independent factors take increasing importance in NTG Two previous meta-analyses have assessed the association between MYOC polymorphisms and POAG susceptibility Although we tried to conduct a thorough review of the existing literature, this study has several potential limitations. First, the possibility of selection biases cannot be completely excluded because all of the included studies were observational, and the potential confounding effect of age and sex might make the interpretation of the results and stratified analyses difficult. Second, only five POAG mutations were included in this analysis. Other potential polymorphisms, such as those at G12R, T123T, D208E, T285T, I288I, T325T, K398R, and A488A, were not included. Third, only published studies were included. Although multiple databases and websites were searched, unfortunately, it is possible that we may have failed to include some papers, especially those published in other languages. We can't find any evidence of publication bias by funnel plots, however, considerable between-study heterogeneity was found for R76K.In conclusion, this systematic review summarized the strong evidence for an association between myocilin polymorphisms and POAG. Our results suggested Q368X and T353I variants of myocilin gene can be taken as reference loci for exploring POAG susceptibility, both in high-tension glaucoma. Furthermore, the prevalence of the two mutations of myocilin gene might be ethnicity-dependent, namely, in Caucasians for Q368X and in Asians for T353I.myocilin, MYOC, trabecular meshwork-induced glucocorticoid response protein, TIGR, GLC1A) and terms related to disease . Additional studies were also identified by a hand search of all the references of retrieved articles.Studies addressing the association between MYOC mutations and polymorphisms and POAG were identified by searching for articles in the PubMed, and EMBASE until 31 December 2011. A broad search strategy combined terms related to gene  case-control study; (2) patients had to be POAG; and (3) Only the most widely mutations and polymorphisms were considered: R46X, R76K, Y347Y, T353I, Q368X. Exclusion criteria were: 1) studies with family-based designs; 2) studies on other polymorphisms other than the target polymorphisms.Data extraction was performed by two reviewers independently and in duplicate. For each study, the following data were extracted: first authors and publication year, country of origin, study base, study participant ethnicity, numbers of cases and controls, diagnosis criteria, demographic data, and genotype distributions for each polymorphism among cases and controls.2 test. If no heterogeneity detected (P>0.1), a fixed effects model was selected to pool the data. A random-effect model, otherwise, was employed after exploring the causes of heterogeneity. Stratified analyses were conducted with respect to ethnicity  and diagnosis criteria . Begg's rank correlation method and Egger's weighted regression method were used to statistically assess publication bias.The association between MYOC polymorphism and POAG was calculated using co-dominant model. We used the odds ratio (OR) and corresponding 95% confidence intervals (CI) as the metric of choice. The statistical analysis was performed by Comprehensive Meta-Analysis . The between-study heterogeneity was tested by the Q test and I


The genetic origins of altruism, defined here as a costly act aimed to benefit non-kin individuals, have not been examined in young children. However, previous findings concerning adults pointed at the arginine vasopressin receptor 1A (AVPR1A) gene as a possible candidate. AVPR1A has been associated with a range of behaviors including aggressive, affiliative and altruistic phenotypes, and recently a specific allele (327 bp) of one of its promoter region polymorphisms (RS3) has been singled out in particular. We modeled altruistic behavior in preschoolers using a laboratory-based economic paradigm, a modified dictator game (DG), and tested for association between DG allocations and the RS3 “target allele.” Using both population and family-based analyses we show a significant link between lower allocations and the RS3 “target allele,” associating it, for the first time, with a lower proclivity toward altruistic behavior in children. This finding helps further the understanding of the intricate mechanisms underlying early altruistic behavior. Altruism, defined here as costly acts aimed at benefiting strangers, is a subgroup of prosocial behaviors . Across mammal species, AVP serves both as a peripheral hormone in regulating water balance as well as a hormone in the brain associated with social behaviors. The role of this gene has been reviewed in both animal 25, RS1 and RS3; CACACACACA). They can affect processes such as translation and transcription The AVPR1A gene is characterized by three promoter region microsatellites ((GT)A growing body of research associated the AVPR1A RS3 327 bp allele  with a range of social skills in both nonclinical and clinical groups Ethics approval for this study was obtained from the ethics committee at Herzog Hospital, Jerusalem, and by the Israeli Ministry of Health higher committee on human medical research. Mothers provided written informed consent before enrolling in the study.mean age = 44.66 months, SD = 3.01). They were part of a larger sample of twins, participating in the Longitudinal Israeli Study of Twins, whose focus is on children's social development as influenced by genetic and environmental effects Participants were three and a half-year old twins,  and extracted with use of the MasterPure kit . Amplification of the RS3 AVPR1A microsatellites was achieved using the following primers 2. ReddyMix buffer consisted of 75 mM Tris-HCl (pH 8.8 at 25°C), 20 mM (NH4)2SO4, and 0.01% (v/v) Tween 20. The sample was initially heated at 95°C for 5 min followed by 30 cycles of 95°C (30 s), 55°C (30 s), and 72°C (40 s), and a final extension step of 72°C for 10 min. The PCR product was analyzed on an ABI 310 DNA analyzer .Each reaction mixture contained 0.5 µM primer and 20 ng of DNA. A ReddyMix master mix (Thermoprime plus DNA polymerase) was used  at a magnesium concentration of 1.5–2.5 mM MgClThe observed length of the RS3 promoter region depends on the PCR primers employed, and the 327 bp allele in the current study corresponds to the 334 allele in other studies http://www.sph.umich.edu/csg/abecasis/PedStats/). SPSS v. 16 (windows) was used for the population-based analysis. In a population-based design the frequencies of the alleles of interest are compared between populations of cases and controls, and a higher frequency in cases is taken as evidence that the allele is associated with the targeted phenotype Mendelian inheritance and Hardy-Weinberg equilibrium were verified with PEDSTATS version 0.6.12 http://www.mrc-bsu.cam.ac.uk/personal/frank/software/unphased/; In order to avoid the confounding effects of population stratification and further validate the findings, the transmission disequilibrium test (TDT) was used for family-based analysis. In the simplest family-based design which uses trios, affected offspring and two parents , the null hypothesis expects transmission according to Mendelian inheritance – all alleles having a 50% chance of being inherited to the next generation. A deviation from the expected allelic transmission indicates an association between the targeted alleles and the phenotype p = 0.4), therefore sex differences were not considered further.There were no significant sex differences in allocations (t(df = 96) = 0.834, The distribution of the number of sticker charts allocated is presented in p = .004), with carriers of the 327 bp allele being 4 times less likely to donate more than one sticker chart . Kendall's tau-c was used due to the ordinal nature of the dependent variable. er chart .A family-based analysis was used in order to further examine the association between the ‘target allele’ and allocations. An important characteristic of family-based designs is their robustness against population substructure 2 = 3.89, p = 0.04, indicating carriers as less likely to allocate sticker charts.As expected from the population based analysis, a significant association was found between the RS3 327 bp repeat, coded as ‘target allele’ vs. all others, and sticker chart allocations, χYoung children's prosocial behavior is now increasingly understood as developing earlier than previously thought The link between AVPR1A and social behavior in general, and altruism in particular, may be partially explained by the association found in adults between the length of RS3 and AVPR1A mRNA expression in the hippocampus The new found link between the AVPR1A RS3 327 allele and children's lower allocations in the DG is in line with previous research that has also examined the RS3 ‘target allele’ effect individually and linked it with lower levels of social skills. Walum, et al. The current finding partly replicates our previous finding in adults, which linked AVPR1A to altruism The main strength of the current study lies in the use of experimental data, specifically, an economic game which is applicable to various age groups and is being widely used in behavioral research. The use of families is also of value. It allowed the addition of a family-based analysis to the population-based analysis, and thus enabled the reliance on allelic transmission instead of solely on group comparisons. Another strength lies in the examination of preschoolers, which enabled the first finding of a main genetic effect on altruistic behavior in such an early age, that is not contingent upon possible reciprocation.The main limitation of this study is the relatively small sample size. However, since the targeting of the AVPR1A gene was based on previous research linking it to social behavior in various samples p = 0.20). Nonetheless, future research should investigate the association between the RS3 ‘target allele’ and preschoolers' altruism in a more private setting.Similar to previous studies with children An accumulating body of research shows AVP's role in social and affiliative behaviors across a diversity of species


TNF-α gene, −308G/A and −238C/T, have been suggested to alter the risk for prostate cancer, but the results have been inconclusive so far. In order to obtain a better understanding of the effects of these two polymorphisms on prostate cancer risk, all available studies were considered in a meta-analysis.Tumor necrosis factor-alpha (TNF-α) is an important inflammatory cytokine that may play a role in controlling the progression of prostate cancer. Two common polymorphisms in the We conducted a comprehensive literature search in the Cochrane Library, PubMed, EMBASE, Chinese Biomedical Literature database (CBM), and the China National Knowledge Infrastructure (CNKI). The associations were evaluated by calculating the pooled odds ratio (OR) with 95% confidence interval (95% CI).In this meta-analysis, we included 14 studies with 5,757 patients and 6,137 control subjects for the TNF-α-308G/A polymorphism and 1,967 patients and 2,004 control subjects for the TNF-α-238C/T polymorphism. A significantly increased prostate cancer risk was found to be associated with the TNF-α-308C/T polymorphism in studies with healthy volunteers . No significant association was found between the TNF-α-238G/A polymorphism and prostate cancer risk in the overall or subgroup analyses. There was no risk of publication bias in this meta-analysis.Our results suggest that while the TNF-α-238G/A polymorphism may not be associated with prostate cancer the TNF-α-308C/T polymorphism may significantly contribute to prostate cancer susceptibility in healthy volunteers.http://www.diagnosticpathology.diagnomx.eu/vs/1629288120116301The virtual slide(s) for this article can be found here:  Prostate cancer is the most common malignant tumour of the male reproductive system in the Western hemisphere. Approximately 238,590 new cases and 29,270 deaths of prostate cancer are reported annually. TherefoTumour necrosis factor-alpha (TNF-α) is a mediator of the inflammatory process that is secreted by monocytes, macrophages, neutrophils, T cells, and NK cells after stimulation. TNF-α is a pro-inflammatory molecule that may play an important role in the development of the immune response,8 and afA number of studies have reported the association between prostate cancer risk and TNF-α promoter polymorphisms –308G/A and/ or –238G/A polymorphisms, but the results are controversial-16. A siWe conducted a comprehensive literature search in the Cochrane Library, PubMed, EMBASE, Chinese Biomedical Literature database (CBM), and the China National Knowledge Infrastructure (CNKI) using the search terms “prostate cancer OR prostate carcinoma OR PCa”, “polymorphism or variant OR mutation OR genotype” and “Tumour Necrosis Factor-alpha OR TNF OR TNF-α” and various combinations of these terms. All the articles were updated on September 10, 2013. The search was performed without any limitations of language. Review articles, original articles, and other studies of interest were examined to identify additional eligible studies.Studies included in the meta-analysis were required to meet the following criteria: (1) studies investigating the association between TNF-α gene polymorphisms and prostate cancer risk; (2) case–control or cohort studies; and (3) the papers must list the sample size, distribution of genotype, and allele frequency. If serial studies of the same population from the same group were reported, the most recent or largest population was chosen. When a study reported the results on different subpopulations, we treated it as separate studies in the meta-analysis. Studies were excluded from our meta-analysis if they met one of the following exclusion criteria: (1) the study was conducted on animals, (2) the design was based on family or sibling pairs, or (3) insufficient original data was available for data extraction, for instance, the number of genotypes could not be ascertained.T Two separate investigators (Ma and Zhao) reviewed and extracted data from the studies included independently to ensure the accuracy of the data. The following parameters were collected from eligible studies: first author's surname, year of publication, country of study population, ethnicity, genotyping methods, sample size, source of controls , matching variables, prostate cancer diagnosis, QC when genotyping, and the number of genotype frequencies in cases and controls. The two investigators verified data accuracy by comparing collection forms between investigators. If different results were generated, they would carry out discussions until a consensus was reached.The quality of the selected studies was independently evaluated by two reviewers . The criteria for quality appraisal are listed in Table 2 metric. I2 values of 75%, 50%, and 25% were considered to reflect high, moderate, and low heterogeneity, respectively[2 < 50% or P > 0.10), the ORs and 95% CI would be estimated for each study in the fixed-effect model (Mantel-Haenszel method)[Crude odds ratios (ORs) and corresponding 95% confidence intervals (CIs) were employed to estimate the strength of association between the two polymorphisms and prostate cancer risk. The recessive genetic model, dominant genetic model and additive genetic models were used to calculate the pooled ORs and 95% CIs for both the polymorphisms. Heterogeneity among the studies included in the meta-analysis was evaluated by the chi-square-based Q test and quantified by the Iectively. When no method). Otherwi method). Logistit test was used to determine the significance of the asymmetry, and if the P value was <0.10, indicating the presence of publication bias, the non-parametric “trim and fill” method was used to adjust for it[A funnel plot was used to verify potential publication bias using the standard error of log (OR) for each publication plotted against its log (OR), and the asymmetry of the funnel plot was tested by Egger’s regression. The t tt for it. SensitiAs shown in Figure 2 = 0.0%, and PQ = 0.602 for heterogeneity) and dominant model AA + AG vs. GG  but not in the recessive model AA vs. AG + GG . Furthermore, excluding the studies by OH et al.[The data suggested no significant association between the TNF-α-308G/A polymorphism and prostate cancer risk in all genetic models  in the overall populations. We further employed meta-regression and subgroup analyses to explore the source of heterogeneity. However, meta-regression analysis of data showed that ethnicity, genotyping methods, source of controls, QC when genotyping, prostatic cancer diagnosis, and quality scores did not affect modifiers. Subsequently, we stratified the studies by ethnicity and source of controls, and observed the lack of homogeneity among Caucasians and hospital-based studies in the two genetic models , Caucasians , and hospital-based studies . We also performed Galbraith plots analysis to identify the outliers that might contribute to the heterogeneity. The study of Oh et al. was an oFor the TNF-α-238G/A polymorphism, substantial heterogeneities were observed between studies in the additive model AG vs. GG (p = 0.014) and dominant model AA + AG vs. GG (p = 0.041) in the overall populations. We further employed subgroup analyses stratified by ethnicity and source of controls. However, heterogeneities were still observed among hospital-based studies in these two genetic models . When we excluded HWE-violating studies, the corresponding pooled ORs were not materially altered, indicating that the data in this meta-analysis are relatively stable and credible.Begg’s funnel plot and Egger's linear regression tests were created to estimate the publication bias risk in this meta-analysis. As shown in Figure TNF-α is a member of the TNF/TNFR cytokine superfamily, and is an intercellular communicating molecule involved in building transient or long-lasting multicellular structures. It is aTwo variations in the promoter region of the TNF-α gene, namely, TNF-α-308G/A and TNF-α-238G/A, have been commonly studied. The G to A substitution at position −308 in the TNF-a promoter increases TNF-α transcription activity and the serum TNF-α level. Indeed,Our meta-analysis summarized for the first time all the available data on the association between TNF-α-238G/A polymorphism and prostate cancer risk, including a total of five studies, involving 1,967 prostate cancer cases and 2,004 controls. Our results demonstrated that the TNF-α-238G/A polymorphism was not significantly associated with prostate cancer risk not only in the overall population but also in the subgroup analyses stratified by ethnicity and source of controls.With respect to TNF-α-308G/A polymorphism, 14 studies including 5,757 prostate cancer cases and 6,137 controls were found in our meta-analysis. The data suggested no significant association between TNF-α-308G/A polymorphism and prostate cancer risk in all genetic models in the overall populations, which is consistent with the previous findings made by Wang et al. and Wang2 values decreased obviously and PQ values were greater than 0.10 in the overall populations and in Caucasians. In addition, excluding the two studies did not significantly affect the results in the different comparison models in the overall population and subgroup analyses. The results indicated that the two studies might be the major source of the heterogeneity for the 308G/A polymorphism.For the TNF-α-308G/A polymorphism, substantial heterogeneities between studies were observed in the additive model AG vs. GG (p < 0.001) and dominant model AA + AG vs. GG (p < 0.001) in the overall populations. To explore the source of heterogeneity, we employed meta-regression and subgroup analyses. Meta-regression analysis revealed no definite source of heterogeneity. Subgroup analyses by ethnicity showed that heterogeneity existed in Caucasian subjects and hospital-based studies in the additive and dominant models. To further investigate the heterogeneity, Galbraith plot analysis was used to identify the outliers that might contribute to the heterogeneity, and two outliers were found. When they were excluded from the analysis, the ISubstantial heterogeneities between studies were observed in the additive and dominant models for the TNF-α-238G/A polymorphism in the Caucasian population and hospital-based studies. As only five studies on the association between TNF-α-238G/A polymorphism and prostate cancer risk were included in our meta-analysis, meta-regression analysis and Galbraith plots analysis were not performed; therefore, these results require further investigation.This meta-analysis has some limitations that must be considered. First, the overall outcomes were based on individual unadjusted ORs, while a more precise estimation should be adjusted by confounding factors such as smoking status, age, and environmental factors. Second, the sample sizes in this analysis were not adequate, especially the African-American populations; therefore, more subjects of different ethnicities would be required to accurately clarify whether ethnicity has a biological influence on cancer susceptibility. Third, the controls were not consistently screened across the studies analysed. Therefore, the control groups may have different risks of developing prostate cancer. Fourth, as only certain published studies were included in our meta-analysis, publication bias is very likely to occur although it was not shown in the statistical test.In conclusion, the present meta-analysis suggests that the TNF-α-238/A polymorphism is unlikely to be a risk factor for prostate cancer, while the TNF-α-308 G/A polymorphism may make a significant contribution to the risk of prostate cancer in healthy volunteers. However, to clarify the role of TNF-α-308G/A and TNF-α-238G/A polymorphism in prostate carcinogenesis, more studies with large samples are needed in the future.TNF-α: Tumor necrosis factor-alpha; SNPs: Single nucleotide polymorphisms; HWE: Hardy–Weinberg equilibrium; OR: Odds ratio; CI: Confidence interval; PCR-RFLP PCR: Based restriction fragment length polymorphism; QC: Quality control.The authors declare that they have no competing interests.All authors have read and approved the final files for this manuscript.


A) receptor, a predominant receptor evoking potent vasoconstrictive action of endothelin-1, contributes to susceptibility to IR-associated hypertension in 1694 subjects of Chinese and Japanese origins. Blood pressures (BPs) and biochemistries were measured. Fasting insulin level, insulin-resistance homeostasis model assessment (HOMAIR) score, and area under curve of insulin concentration (AUCINS) were selected for assessing insulin sensitivity. Genotypes were obtained by methods of polymerase chain reaction-restriction fragment length polymorphism. Foremost findings were that minor allele frequency of the T323C polymorphism was noticeable lower in our overall Asian subjects compared to multi-national population reported in gene database; moreover both the genotypic and allelic frequencies of the polymorphism were significantly different between the two ethnic groups we studied. The genotype distributions at TT/TC/CC were 65, 31, 4% in Chinese and 51, 41, 8% in Japanese, respectively (p < 0.0001). Additionally, carriers of the C homozygote revealed characteristics of IR, namely significantly higher levels of fasting insulin, HOMAIR score, and AUCINS at 29.3, 35.3, and 39.3%, respectively, when compared to their counterparts with TT/TC genotypes in Chinese. Meanwhile, the CC genotype was associated with a higher level of high density lipoprotein cholesterol in Japanese. No association of the polymorphism with BP was observed. This study demonstrated for the first time that T323C polymorphism of ETA receptor gene was associated with an adverse insulin response in Chinese and a favorite atherogenic index in Japanese.Co-heritability of hypertension and insulin resistance (IR) within families not only implies genetic susceptibility may be responsible for these complex traits but also suggests a rational that biological candidate genes for hypertension may serve as markers for features of the metabolic syndrome (MetS). Thus we determined whether the T323C polymorphism (rs5333) of endothelin type A (ET Insulin resistance (IR) and hypertension are related traits. Individuals who are insulin resistant are glucose intolerant, hyperinsulinemic, have higher blood pressure (BP) as well as atherogenic dyslipidemia . The cluA) and endothelin type B receptor (ETB) which bind the three endothelins with various affinities  was associated with IR and hyperinsulinemia in Chinese, and was related with high density lipoprotein (HDL)-cholesterol and atherogenic index in Japanese.The present study was investigated to address the genetic polymorphism of ETSubjects in this current study were those for the Stanford Asian-Pacific Program in Hypertension and Insulin Resistance (SAPPHIRe) genetic study that aimed at mapping genetic loci for hypertension and IR in a well-characterized population regarding IR and its related metabolic traits. The study design and subject recruitment for SAPPHIRe were described in detail previously . In brieIR), a surrogate index for insulin sensitivity validated to the reference method of the euglycemic hyperinsulinemic clamp , were selected for assessing insulin sensitivity. Clinical chemistries included fasting concentrations on total cholesterol, triglycerides, HDL cholesterol were determined by automated enzymatic methods using colorimetry as end point. Low density lipoprotein (LDL)- and very low density lipoprotein (VLDL)-cholesterol were calculated by Friedewald equation , and administration of an informative questionnaire to obtain biochemical data and phenotypic information on subjects. Measurement protocols for BP and anthropometry were previously reported in detail , 32. Bodic clamp , using tic clamp : fastingequation . All lipA receptor gene. The DNA fragment containing the polymorphism site was amplified using the forward primer 5′-TTTCTCACTTTCCTTTAGCG-3′ and the reverse primer 5′-ACCTAAGTAATTCACATCGG-3′. PCR amplifications were carried out in a final volume of 30 μL by 30 cycles at 94, 50, and 72°C each for 30 s and followed by a final extension step at 72°C for 10 min. Blank controls containing no genomic DNA were included in each run of PCR amplification to monitor contamination. Agarose gel electrophoresis on randomly selected PCR products was performed routinely to check if amplification succeeded. For subsequent RFLP analysis, the PCR product was mixed with 1U AflII and 1 μg/mL BSA in the appropriate buffer  and incubated at 37°C for 3 h to ensure the complete enzyme digestion. The digested products were separated by electrophoresis on 3% agarose gel, visualized by ethidium bromide staining and analyzed under UV light. The person who assessed the genotype was blinded to the clinical data of the subjects from whom the samples originated. Direct DNA sequencing using ABI PRISM377 machine together with BigDye™ Terminator Cycle Sequencing Ready Reaction Kit  on randomly selected PCR products from each genotype was also performed to validate the polymorphisms.Using polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) analysis, we determined the polymorphism of exon 6 codon 323 T → C in the ETAflII is specific for the sequence C/TTAAG. Digestion of PCR product with AflII resulted in two fragments of different sizes at 67 and 87 bp for the existence of C allele. However, the restriction site is lost in the presence of T variant, and therefore digestion products shown a single band at 154 bp. Thus, three genotypes TT, TC, and CC at codon 323 in exon 6 of ETA receptor gene were defined according to numbers of fragments by RFLP, depending on the presence of C allele as a restriction site for AflII.The restriction enzyme A receptor genotype distributions and allele frequencies between subjects with different ethnicity and BP status were compared by Chi-square analyses. The Hardy–Weinberg equilibrium was also examined using the Chi-square test. The association of the ETA receptor gene variant with the variables of interest was assessed using mixed model regression analyses as described previously , and tests were two-sided throughout and a p-value of <0.05 was considered statistically significant.Data were expressed as the mean and the standard deviation (mean ± SD), unless otherwise specified. The ETeviously , with faeviously , 36. SpeThe general characteristics of the participants categorized by ethnicity are shown in Table p = 0.166 for overall, p = 0.225 for Chinese, and p = 0.949 for Japanese). Significant differences (p < 0.0001) on genotype frequencies were observed between subjects with different ethnic backgrounds . Neither genotype nor allele frequency was different with respect to gender. Moreover, there was no statistical significant difference in genotypes concerning the BP status in our studied subjects despite the anticipated role of ETA receptor in the regulation of cardiovascular complications.A total of 1694 biethnic sibling subjects (1304 Chinese and 390 Japanese) deriving from eligible families were analyzed. The genotype distributions in overall or each for Chinese and Japanese were all in compliance with the Hardy–Weinberg equilibrium .Due to the significant ethnic differences, we next evaluated the association of genotypes of ETes Table . In the ol Table . The adjA receptor genotype, BMI, age, et cetera as independent variables. By this approach we could predict if any attribute, while controlling for other covariates, is independently associated with changes in the indices at all. Results on the levels of significance in the pooled studied subjects are present in Table A receptor T323C genotype was not only significantly (p = 0.0012) but also independently from BMI, waist circumference, triglycerides and HDL-cholesterol concentrations, and gender correlated with the index of AUCINS. Additionally, we observed that waist circumference turned into a more significant predictor than WHR to HOMAIR and AUCINS in our studied population.Since phenotypic expression of IR-associated hypertension is diverse and bounded, we further performed correlation analysis by treating the indices of insulin sensitivity as dependent variables, and the potential contributors including the ETA receptor, and an amelioration of hypertension via administration of selective ETA receptor antagonist observed previously in our fructose-induced hypertensive rats  at 0.22 and 0.35, respectively. Secondly, an ethnic difference between Chinese and Japanese we studied on the allele frequencies of the T323C variant was identified. Thirdly, the ETA receptor T323C polymorphism was associated with discrete features of the MetS in our two Asian populations.Enhanced vascular gene expressions of ET-1 and ETive rats  and in cive rats , 37, 38 ion gene , 29, 39 ion gene , 40. TheA receptor gene were observed between our subjects of Chinese and Japanese origin, no association of the T323C polymorphism with hypertension status was seen in either Asian ethnic group, i.e., neither systolic nor diastolic BP appeared to be influenced by the gene variant. Our findings supported and extended the prior observations in Caucasians . As a silent polymorphism which nucleotide substitution on the third base of the codon does not alter the amino acid sequence of the receptor protein, it is unlikely that the variant itself is involved in the pathophysiology of MetS. One possibility for this silent polymorphism exerts its influence is that it is in linkage disequilibrium with a nearby causal variant which has not yet been identified. In addition, the wobble base pair is a fundamental unit for RNA secondary and tertiary structures and therefore may have functional consequences. Without further RNA studies, the possibility of unrecognized alleles, disrupting exonic splicing enhancers, and/or influencing gene transcription cannot be excluded. Although BP and IR have been shown to cosegregate in Hispanic families with a hypertensive proband (Human genome contains a single copy of the ETrtension . It shou proband .A1/A2) with stroke in a sub-group of high risk hypertension patients (Beyond candidate genes in ETS for the predisposition of IR-associated hypertension, the notion of using biological candidate genes for interrelating traits as genetic markers for the features of MetS and/or diabetes has also been applied in other studies regardless successes or not. For example, Bayoglu et al. identified polymorphism rs10757278 on chromosome 9p21 which is risk locus for coronary artery disease may confer increased risk for MetS in Turkish people . Ned et patients . Moreovepatients , 53 and patients , or for patients , 55 and patients . Given tpatients .A receptor gene to features of MetS in this study though no association between the gene variant and BP is reassuring in our hypertensive subjects of Chinese and Japanese origin. Our findings can be considered as the first time to be related directly to IR, and also the first in Asian populations despite its dissociated effects on different ethnicities. If reproducible in other independent ethnicities, our results reported herein suggest that the polymorphism of ETA receptor gene may serve as one of the independent determinants for manifested IR which is often a prelude to hypertension. Nevertheless, the functional significance of this particular gene variant, the inference on haplotype effects, and the gene–gene/gene-environment interactions remain to be elucidated. Determinants of CVD risk in diabetes beyond hyperglycemia has been an important issue to address lately, since intensive control of plasma sugar alone is often insufficient to reduce the incidence of major CVD in these patients (In conclusion, we have successfully related the T323C polymorphism of ETpatients . Componepatients .Guarantor of integrity of the study and study concepts: Low-Tone Ho. Definition of intellectual content: Low-Tone Ho, John Grove, Thomas Quertermous. Literature research: Yung-Pei Hsu, Chi-Chung Juan, Low-Tone Ho. Clinical studies: Low-Tone Ho, Chih-Tai Ting, Kuang-Chung Shih, Lee-Ming Chuang, Kamal Masaki. Genotyping experiments: Yung-Pei Hsu, Chi-Chung Juan. Data acquisition: Chin-Fu Hsiao, Ming-Wei Lin, Yung-Pei Hsu, Chi-Chung Juan. Statistical analysis: Chin-Fu Hsiao, Ming-Wei Lin, Shu-Chiung Chiang. Manuscript preparation: Yung-Pei Hsu, Low-Tone Ho. Manuscript editing and revise: Low-Tone Ho, Yii-Der I. Chen.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Power analysis was performed with the Power and Sample Size Calculation (PS) program. A fixed or random effect model was used on the basis of heterogeneity. Publication bias was quantified and examined with the Begg's funnel plot test and Egger's linear regression test. The meta-analysis was performed with Review Manager 5.1 and Stata 11.0.ORs (95% CIs) for TNF-α −308G>A of A vs. G allele and GA+AA vs. GG genotype were 1.63 (1.17–2.25) and 1.47 (1.17–1.85), respectively.There were 10 studies including 1425 T2DM patients and 1116 healthy control subjects involved in this meta-analysis. No significant publication bias was found in the studies. The pooled α −308G>A polymorphism was strongly associated with T2DM risk, and A allele at this locus might be a susceptibility allele for the development of T2DM in Han Chinese population.This meta-analysis result suggested that TNF- It has been shown that TNF-α can affect insulin resistance by regulating adipocyte gene expression Diabetes mellitus (DM) has become a major health problem in the world. It is estimated that the population with DM will rise to 300 million in the year of 2025, and most of them are type 2 diabetes mellitus (T2DM) α polymorphism and T2DM risk have been published in different countries. However, the findings from these studies are largely inconsistent. Particularly in China, the obvious contradictory results across different studies were obtained, rendering interpretation of its genetic risk at large for Han Chinese people difficult and confusing. Most studies of Han Chinese population focused on the association of TNF-α −308G>A, a genetic variant in the promoter region of this gene, with T2DM risk, because it may play an important regulatory role for TNF-α production. The schematic diagram of TNF-α −308G>A together with the location of the candidate SNP rs1800629 was shown in So far, a number of studies α polymorphisms become essential. The aim of this meta-analysis was to integrate the findings from multiple studies to provide an overall assessment whether TNF-α −308G>A (rs1800629) polymorphism is associated with T2DM risk in a population of Han Chinese subjects.Nevertheless, the association between this polymorphism and T2DM susceptibility still remained unclear in Han Chinese population, with inconsistent findings across different studies. Since each single study was limited in providing sufficient information and adequate power for Han Chinese population, summarizing the results from these individual studies for obtaining a reliable and consolidated conclusion regarding TNF-http://www.cnki.net/), Chinese VIP database (http://www.cqvip.com/), Chinese Wanfang database (http://g.wanfangdata.com.cn/) and MEDLINE database (http://www.ncbi.nlm.nih.gov/pubmed), up to September 2012, for all publications about the association between TNF-α −308G>A polymorphism and T2DM in Han Chinese population. The keywords in this search were consisted of TNF-α, type 2 diabetes mellitus, polymorphism and Chinese/China.Major electronic literature databases were systematically searched, which included China National Knowledge Infrastructure (CNKI) database  P-values of HWE test in healthy control groups. Then, a group discussion was conducted to remedy any discrepancies in data collected by different reviewers.Two reviewers independently extracted data from relevant studies, including author, year of publication, geographic region, genotype distribution for T2DM patients and healthy control subjects, descriptive statistics on each study  and http://biostat.mc.vanderbilt.edu/wiki/Main/PowerSampleSize) χ2-based Q statistic P<0.05), a random effect model (Dersimonian-Laird method) was selected to merge data. Otherwise, a fixed effect model (Mantel-Haenszel method) was employed to analyze data. Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated to assess the association between −308G>A of TNF-α and T2DM risk. Then, publication bias was estimated by Begg's funnel plot test and Egger's linear regression test. Finally, sensitivity analysis was performed to assess the influence of each individual study.This meta-analysis was performed with Review Manager 5.1 and Stata 11.0. Power analysis was performed using the Power and Sample Size Calculation (PS) program (α −308G>A (rs1800629) polymorphism and T2DM susceptibility in Han Chinese were identified. Then, one study P<0.05). Hence, there were actually 10 studies At first, 11 studies of the association between TNF-Before this meta-analysis, power analysis was performed by using the PS program to prove whether the selected studies could offer adequate power. With data provided by the 10 studies, Mantel-Haenszel test, assuming that each 2×2 table consists of cases and controls selected from a different stratum that is defined by studies, could provide the power of 0.996 and 0.916 under the allelic and dominant genetic models, respectively.P = 0.03). Therefore, for estimating the overall allelic risk, a random effect model was selected to merge data. ORs (95% CIs) estimate of TNF-α −308G>A  for each study and the overall estimate for the pooled data. The overall allelic OR (95% CI) was 1.63 (1.17–2.25), indicating that this allele brought a modest-size allelic risk of diabetes to Han Chinese population.There was statistically significant allelic heterogeneity among the included studies . Hence, a fixed effect model was employed to analyze data. The pooled OR (95% CI) for TNF-α −308G>A under the dominant model (GA+AA vs. GG genotype) was 1.47 (1.17–1.85) (Association between genotypes (GA+AA vs. GG genotype) at TNF-17–1.85) , also inα −308G>A for A vs. G allele and GA+AA vs. GG genotype were shown in P-values for Begg's test of TNF-α −308G>A A vs. G allele and GA+AA vs. GG genotype were 0.531 and 0.180, respectively, and the P-values for Egger's linear regression test were 0.323 and 0.123, respectively  to 1.72 (1.19–2.48) for A vs. G allele, and 1.37 (1.08–1.75) to 1.61 (1.24–2.10) for GA+AA vs. GG genotype, respectively, suggesting that the pooled estimates of allelic and genotypic risks obtained in this study were both stable and reliable.Finally, a sensitivity analysis was performed to evaluate the influence of individual studies. A leave-one-out procedure  was used to estimate the contribution of a single study to the pooled α gene, situated on human chromosome 6p21.3, encodes a multifunctional proinflammatory cytokine, which plays the most important role in the development of T2DM. It has been well documented that high expression of TNF-α in adipose tissue is related to insulin resistance, considered to be an important pathogenic mechanism for the development of T2DM TNF-α −308G>A (rs1800629) and T2DM is a hot focus. By searching literature, we found that the findings regarding this association were of obvious discrepancies across different studies. Hence, it is highly demanded to systematically assess the association of TNF-α −308G>A with T2DM prior to translation of this genetic finding into clinical practice.Recently, a number of candidate-gene studies for diabetes in Han Chinese population have been reported, and the association between TNF-α −308G>A and T2DM in Han Chinese population, and the pooled data from 10 eligible studies contained 1425 T2DM patients and 1116 healthy subjects. This meta-analysis showed that there was statistically significant association between TNF-α −308G>A polymorphism and T2DM risk in Han Chinese population, demonstrating that TNF-α −308G>A was an important risk factor for T2DM in Han Chinese population. This conclusion is largely consistent with a previous meta-analysis examining this association in Asian populations α −308G>A gene polymorphism was not associated with T2DM risk in these large racial divisions. These discrepancies suggest that the genetic effect of TNF-α −308G>A is race-specific. While it imposes a significant risk on Han Chinese population, TNF-α −308G>A polymorphism might not be associated with T2DM risk in other racial populations such as Caucasians To the best of our knowledge, this study was the first meta-analysis to comprehensively examine the association between TNF-To avoid potential racial admixture, we restricted our analysis to Han Chinese in this study. All of the 10 included studies were of Han Chinese ancestry. Nevertheless, for Han Chinese population who live in a vast country, geographical and culture differences might be additional factors causing genetic heterogeneity. In this meta-analysis, five of the included studies were from northern China, and the remaining five from southern China. The culture and habits, such as personality, diet, living environment, customs and so on, in these two large geographic areas, were very different, and the genetic communications between these two geographic areas might not be as frequently as within each geographic area, which may lead to population stratification. The fact that significant between-study heterogeneity in allelic association and marginal significance in genotypic association across the 10 studies might support our perspective. Other factors  might contribute to heterogeneity as well. Unfortunately, due to limited number of studies and lack of size of samples, we did not perform any stratified analysis regarding these potential confounding factors.α gene with various environmental factors. Finally, only single SNP (rs1800629) in TNF-α gene was analyzed in this study, and whether additional genetic variants of either functional or polymorphic contribute to this gene remains unclear, which may lead to underestimation of its overall genetic effect on T2DM susceptibility. Thus, constructing its spectrum of genetic variants is required for comprehensive assessment of its role in conferring susceptibility to T2DM.Finally, we should recognize several limitations of this study. First, we did not take into account the impact of several covariates  on the results of this meta-analysis due to incomplete data for several studies. Second, because of lack of the data for individuals’ environmental exposures, we did not further explore the interactions of TNF-α −308G>A (rs1800629) polymorphism were statistically significantly associated with T2DM in Han Chinese population, suggesting that A allele at this locus increases the risk for the development of T2DM in this population. However, in our view, more large-scale independent studies or larger sample sizes are still needed to consolidate this finding in the future. Furthermore, for a complex disease like T2DM, which is considered being the result of the sophisticated interplays between multiple genes, and their interactions with environment, a rational and extended research is to further identify the genes and environmental triggers interacting with TNF-α −308G>A and the functional involvement of this polymorphism leading to the molecular pathogenesis of T2DM.In conclusion, by systematically integrating 10 well-characterized association studies for Han Chinese, with 1425 T2DM patients and 1116 healthy subjects, we found that TNF-Figure S1PRISMA flow diagram of including studies.(DOC)Click here for additional data file.Checklist S1PRISMA checklist of the association between TNF-α −308G>A and T2DM.(DOC)Click here for additional data file.


To assess the association of P2RX7 gene rs2230912 polymorphism with mood disorders using a meta-analysis.Data were collected from the following electronic databases: PubMed, Excerpta Medica Database, Elsevier Science Direct, Cochrane Library, and Chinese Biomedical Literature Database, with the last report up to April 1, 2013. Odds ratio (OR) with 95% confidence interval (CI) was used to assess the strength of the association. Dependent on the results of heterogeneity test among individual studies, the fixed effect model (Mantel–Haenszel) or random effect model (DerSimonian–Laird) was selected to summarize the pooled OR.P>0.05). We also performed disease-specific meta-analysis in unipolar depression and bipolar disorder. No significant association of this polymorphism with unipolar depression or bipolar disorder was found (P>0.05). Additionally, we performed subgroup analysis by different types of cases. No significant association of this polymorphism with mood disorders in clinical cohorts or population-based cohorts (P>0.05). A significant association of this polymorphism with mood disorders was found for the allele contrast in family-based cohorts .We identified 13 separate studies using search . We detected significant between-study heterogeneity. No significant association of this polymorphism with mood disorders was found (Overall, our meta-analysis suggests that P2RX7 gene rs2230912 polymorphism may not contribute to the risk of developing mood disorders using a case-control design. Given the discordance in the subgroup analysis by different types of cases, further studies based on larger sample size are still needed. Mood disorders define a large group of human psychiatric disorders with a high phenotypic complexity but all characterized by consistent, pervasive alterations in mood, which affect thoughts, emotions and behaviours, and they are among the most prominent causes of disability as well as the second leading source of disease burden The P2X7 receptor is an ATP-gated non-selective cation channel activated by high concentrations of ATP (>100 µM), expressed as homo-oligomeric assemblies of individual subunits, and is widely distributed at immunocompetent cells of the central and peripheral nervous system P2RX7  gene located on chromosome 12q24.31 encodes the P2X7 receptor The present meta-analysis was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA) guidelines  [27]. WeTwo reviewers independently extracted the data with the standard protocol, and the result was reviewed by a third reviewer. Discrepancies were resolved by discussion with our research team. From each study, we extracted the first author, year of publication, journal, ethnicity, types of mood disorders, numbers of cases and controls, and the available genotype and allele frequencies of P2RX7 gene rs2230912 polymorphism. If original data were unavailable in relevant articles, a request for additional data were sent to the corresponding author.P<0.10) indicated heterogeneity across studies. The degree of heterogeneity was further assessed with the 2I statistics (2I = 100%×(Q-df)/Q) P<0.05 (two-tailed) was considered statistically significant.We assessed the relationship between the allele, as well as genotypes and susceptibility to mood disorders. The odds ratio (OR) and its 95% confidence interval (95%CI) were estimated for each study. The heterogeneity between the study results was assessed by the Chi square-test based Q-statistic The characteristics of studies included in the current meta-analysis are presented in P>0.05). A sensitivity analysis was performed in those studies fulfilling HWE. The result showed that there was still no significant association of P2RX7 gene rs2230912 polymorphism with mood disorders (data not shown).P>0.05). In the analysis of bipolar disorder, significant between-study heterogeneity was found in the contrasts of AG+GG versus AA and AG versus AA. Similarly, no significant association of P2RX7 gene rs2230912 polymorphism with bipolar disorder was found (P>0.05).We also performed disease-specific meta-analysis in unipolar depression  and bipolar disorder . In the analysis of unipolar depression, significant between-study heterogeneity was found in the contrasts of G versus A, AG+GG versus AA and AG versus AA. We did not detect significant association of P2RX7 gene rs2230912 polymorphism with unipolar depression (P>0.05). A significant association of this polymorphism with mood disorders was found for the allele contrast in family-based cohorts .Additionally, we performed subgroup analysis by different types of cases . Significant between-study heterogeneity was found for the contrasts of G versus A, AG+GG versus AA and AG versus AA in clinical cohorts, but not for other contrasts. No significant association of this polymorphism with mood disorders in clinical cohorts or population-based cohorts  may contribute to heterogeneity.Funnel plot and Egger's linear regression test were performed to estimate the publication bias of literatures. The shapes of the funnel plots did not reveal any evidence of obvious asymmetry (funnel plots not shown). The results of Egger's linear regression test are shown in There is compelling evidence from family, twin and adoption studies for the existence of genes influencing susceptibility to mood disorders In the central and peripheral nervous system, the P2X7 receptors are expressed on microglial cells, neurons and astrocytes. They take part in inflammatory responses and the cross-talk between glia and neurons, and are proposed to promote neurotransmitter release at presynaptic sites Several specific details merit consideration in the current meta-analysis. A first consideration is that only published studies were included in this meta-analysis, and Egger's linear regression test was not applied for some comparisons due to the small number of studies. Thus, publication bias may occur. A second consideration is that significant between-study heterogeneity was detected in some comparisons, and may be distorting the results of the meta-analysis. Further studies based on refined phenotypes may help to elucidate the association of P2RX7 gene rs2230912 polymorphism with mood disorders. A third consideration is that our reulsts are based on unadjusted estimates, and a more precise analysis stratified by age and sex could be performed if individual data were available. Finally, all of the eligible studies were conducted in Cauasian population. Further studies are still needed in non-Cauasian population.Overall, our meta-analysis suggests that P2RX7 gene rs2230912 polymorphism may not contribute to the risk of developing mood disorders using a case-control design. Given the discordance in the subgroup analysis by different types of cases, further studies based on larger sample size are still needed.Checklist S1PRISMA checklist.(DOC)Click here for additional data file.


Many studies have examined the association between the CYP1A1 Ile462Val gene polymorphisms and lung cancer risk in various populations, but their results have been inconsistent. To assess this relationship more precisely, a meta-analysis was performed. Ultimately, 43 case-control studies, comprising 19,228 subjects were included. A significantly elevated lung cancer risk was associated with 2 Ile462Val genotype variants  vs Ile/Ile: OR = 1.15, 95% CI = 1.07–1.23) in overall population. In the stratified analysis, a significant association was found in Asians, Caucasians and lung SCC, not lung AC and lung SCLC. Additionally, a significant association was found in smoker population and not found in non-smoker populations. This meta-analysis suggests that the Ile462Val polymorphisms of CYP1A1 correlate with increased lung cancer susceptibility in Asian and Caucasian populations and there is an interaction with smoking status, but these associations vary in different histological types of lung caner. Lung cancer remains the most lethal cancer worldwide, despite improvements in diagnostic and therapeutic techniques Cytochrome P450 1A1 (CYP1A1) metabolizes several suspected procarcinogens, particularly polycyclic aromatic hydrocarbons (PAHs), into highly reactive intermediates An association between CYP1A1 polymorphisms and lung cancer was first reported by Kawajiri and co-workers in 1990 among an Asian study population A single study might not be powered sufficiently to detect a small effect of the polymorphisms on lung cancer, particularly in relatively small sample sizes. Various types of study populations and study designs might also have contributed to these disparate findings. To clarify the effect of the CYP1A1 Ile462Val (exon 7) polymorphism on the risk for lung cancer, we performed an updated meta-analysis of all eligible case-control studies to date and conducted the subgroup analysis by stratification according to the ethnicity source, histological types of lung caner and smoking status of case.The electronic databases PubMed, Embase, Web of Science, and CNKI  were searched for studies to include in this meta-analysis, using the terms “CYP1A1,” “Cytochrome P450 1A1,” “polymorphism,” and “lung cancer.” An upper date limit of March 01, 2012 was applied; we used no lower date limit. The search was performed without any restrictions on language and was focused on studies that had been conducted in humans. We also reviewed the Cochrane Library for relevant articles. The reference lists of reviews and retrieved articles were hand searched simultaneously. When more than one of the same patient population was included in several publications, only the most recent or complete study was used in this meta-analysis.For inclusion, the studies must have met the following criteria: they (1) evaluated CYP1A1 Ile462Val (exon 7) gene polymorphisms and lung cancer risk; (2) were case-control studies or nested-case control study; (3) supplied the number of individual genotypes for the CYP1A1 Ile462Val (exon 7) polymorphisms in lung cancer cases and controls, respectively; and (4) demonstrated that the distribution of genotypes among controls were in Hardy-Weinberg equilibrium.Information was carefully extracted from all eligible publications independently by two authors according to the inclusion criteria. Disagreements were resolved through a discussion between the two authors. The following data were collected from each study: first author’s surname, year of publication, ethnicity, total numbers of cases and controls, and numbers of cases and controls who harbored the Ile462Val (exon 7) genotypes, respectively. We did not contact the author of the primary study to request the information. Ethnicities were categorized as Asian, Caucasian, and mixed. Histological type of lung cancer was divided to lung squamous carcinoma (SCC), adenocarcinoma (AC) and small cell lung cancer (SCLC) in our meta-analysis. The definition of smoking history is very complicated. The smoking histories covered different periods if changes in the number of cigarettes smoked per day or type of tobacco products occurred. According to the general standards, non-smokers were defined as subjects who had smoked less than 100 cigarettes in their lifetime. Although the precise definition of never-smoking status varied slightly among the studies, the smoking status was classified as non-smokers (or never smoker) and smokers (regardless of the extent of smoking) in our meta-analysis. We did not define any minimum number of patients to include a study in our meta-analysis.OR (odds ratios) with 95% CIs were used to determine the strength of association between the CYP1A1 Ile462Val (exon 7) polymorphisms and lung cancer risk. We evaluated this risk with regard to combinations of variants  versus the wild-type homozygotes (Ile/Ile).P value greater than 0.10 for the Q-test indicated a lack of heterogeneity among the studies. Thus, the pooled OR estimate of each study was calculated using the fixed-effects model (the Mantel–Haenszel method) The pooled ORs for the risk were calculated. Subgroup analyses were performed by ethnicity. Heterogeneity assumptions were assessed by chi-square-based Q-test One-way sensitivity analyses were performed to determine the stability of the results–each individual study in the meta-analysis was omitted to reflect the influence of the individual dataset on the pooled OR Potential publication biases were estimated by funnel plot, in which the standard error of log (OR) of each study was plotted against its log (OR). An asymmetrical plot suggests a publication bias. Funnel plot asymmetry was assessed by Egger’s linear regression test, a linear regression approach that measures the funnel plot asymmetry on a natural logarithm scale of the OR. The significance of the intercept was determined by t-test, as suggested by Egger  All calculations were performed using STATA, version 10.0 .Two hundred and fifty-two potentially relevant citations were reviewed, and 43 publications met the inclusion criteria and included in our meta-analysis Of the 43 publications, 35 were published in English and 8 were written in Chinese. The sample sizes ranged from 104 to 1824. All cases were histologically confirmed. The controls were primarily healthy populations and matched for age, ethnicity, and smoking status, 15 studies were hospital-based control and 28 were population-based control. There were 24 groups of Asians, 11 groups of Caucasians, and 8 mixed populations.P = 0.004 for heterogeneity; for Ile/Val and Val/Val combined vs Ile/Ile: OR = 1.15, 95% CI = 1.07–1.23, P<0.001 for heterogeneity)  .P = 0.016 for heterogeneity) and Ile/Val and Val/Val combined vs Ile/Ile . A significant association was also observed in Caucasian carriers of Val/Val vs Ile/Ile  and Ile/Val and Val/Val combined vs Ile/Ile . However, no significant associations were observed in mixed populations for both Val/Val vs Ile/Ile  or Ile/Val and Val/Val combined vs Ile/Ile   .P = 0.004 for heterogeneity) or Ile/Val and Val/Val combined vs Ile/Ile  and Ile/Val and Val/Val combined vs Ile/Ile . However, for non-smokers, no significant associations were observed for both Val/Val vs Ile/Ile  or Ile/Val and Val/Val combined vs Ile/Ile  (Ten out of 40 studies included the association of CYP1A1 exon 7 genotype and lung caner risk stratified by smoking status (non-smokers or never smokers and smokers) . For smogeneity) .A single study involved in the meta-analysis was deleted each time to reﬂect the inﬂuence of the individual data-set to the pooled ORs, and the corresponding pooled ORs were not materially altered (data not shown).Begg’s funnel plot and Egger’s test were performed to identify any publication bias. The funnel plots did not exhibit any patent asymmetry . By EggeCYP genes are large families of endoplasmic and cytosolic enzymes that catalyze the activation and detoxification, respectively, of reactive electrophilic compounds, including many environmental carcinogens . CYP1A1 is a phase I enzyme that regulates the metabolic activation of major classes of tobacco procarcinogens, such as aromatic amines and PAHs When stratified according to ethnicity, a significantly increased risks were identified among Asians and Caucasians for the 2 exon 7 genotype variants, however no significant association was found in mixed population. These findings indicate that polymorphisms of CYP1A1 exon 7 polymorphism may be important in specific ethnicity of lung cancer patients. Population stratification is an area of concern, and can lead to spurious evidence for the association between the marker and disease, suggesting a possible role of ethnic differences in genetic backgrounds and the environment they lived in There are growing biological and epidemiological data to suggest that different lung cancer pathological subtypes, particularly the two most common, are distinct etiological entities that should be analyzed separately As we know, aside from genetic factor, smoking is the major risk factor of lung cancer. Most studies out of 40 studies reported information on smoking habits of cases and controls, however only ten eligible publications provided non-smokers information. Our meta-analysis results showed that a significantly increased risk was found to be associated with the CYP1A1 exon 7 gene polymorphisms and lung cancer risk in smokers, however, no significant association was found among non-smokers. The I-squared value of non-smokers groups is lower than the I-squared values for the smoker population studies, suggesting less heterogeneity among non-smokers populations. Tobacco smoke contains many of carcinogens and procarcinogens, such as benzopyrene and nitrosamine. These compounds are metabolized by the phase I enzymes including CYP family enzymes and converted to inactivemetabolites by the phase II enzymes. Our results should indicate the interaction between CYP1A1 exon 7 gene polymorphisms and smoking in the development of lung carcinoma. However, the association between the extent of smoke exposure and lung caner risk was not clear, further studies with larger sample size are needed to provide insights into the association.Some limitations of this meta-analysis should be acknowledged. First, heterogeneity can interfere with the interpretation of the results of a meta-analysis. Although we minimized this likelihood by performing a careful search of published studies, using explicit criteria for a study’s inclusion and performing strict data extraction and analysis, significant interstudy heterogeneity nevertheless existed in nearly every comparison. The presence of heterogeneity can result from differences in the selection of controls, age distribution, and prevalence of lifestyle factors. Further, only published studies were included in this meta-analysis. The presence of publication bias indicates that non-significant or negative findings might be unpublished. Finally, in the subgroup analyses, different ethnicities were confused with other population, which may bring in some heterogeneity. As studies among the Indians and Africans are currently limited, further studies including a wider spectrum of subjects should be carried to investigate the role of these variants in different populations.In conclusion, the results of our meta-analysis have provided the comprehensive and convincing evidence that CYP1A1 exon 7 polymorphisms are an important modifying factor in determining susceptibility to lung cancer. The effect of CYP1A1 exon 7 gene polymorphisms is diverse by the subgroup analysis stratified by ethnicity, histological types of lung caner and gender of case and control population. More importantly, our study confirms that there is an interaction between two genotypes of CYP1A1 exon 7 gene polymorphisms and smoking. For future studies, strict selection of patients, well-matched controls and larger sample size will be required. Moreover, gene–gene and gene–environment interactions should also be considered.


SLC11A1 gene, has been described to regulate macrophage activation and be associated with infectious and autoimmune diseases. The relation between SLC11A1 polymorphisms and tuberculosis susceptibility has been studied in different populations.Natural resistance associated macrophage protein 1 (NRAMP1), encoded by the SLC11A1 polymorphisms and tuberculosis susceptibility until September 15, 2010 and quantitatively summarized associations of the most widely studied polymorphisms using meta-analysis.We systematically reviewed published studies on SLC11A1 polymorphisms with summarized odds ratio of 1.35 , 1.25 , 1.23 , 1.31  for 3′ UTR, D543N, INT4, and 5′ (GT)n, respectively. Heterogeneity between studies was not pronounced, and the associations did not remarkably vary in the stratified analysis with respect to study population and study base.In total, 36 eligible articles were included in this review. In Meta-analysis, significant associations were observed between tuberculosis risk and widely studied SLC11A1 polymorphisms and tuberculosis susceptibility observed in our analyses supports the hypothesis that NRAMP1 might play an important role in the host defense to the development of tuberculosis.The association between  SLC11A1 gene, has multiple effects on macrophage activation and has been reported to play an important role in host innate immune response against infections Host genetic susceptibility to infectious disease has been widely studied in recent years, which is helpful for high-risk population identification and therefore promotes diseases prevention and early diagnosis Mycobacterium tuberculosis, remains a major challenge to global public health. As estimated, that one-third of the world's population is infected, but that only a minority of those infected ever develop TB SLC11A1 polymorphisms and TB susceptibility was reported in a population from West Africa SLC11A1 loci have been extensively investigated, including 3′ UTR (1729+55del4), D543N (Asp543Asn), INT4 (469+14G/C), and 5′ promoter (GT)n. However, the results were not consistent between the studies. A meta-analysis, which based on literature review until December 2004, suggested an ethnicity specific effect of SLC11A1 polymorphisms on TB risk Tuberculosis (TB), caused by infection of SLC11A1 polymorphisms and TB susceptibility.In this article, we performed a systematic review and meta-analysis, based on literature identification until 15 September 2010, to summarize associations between the most widely studied SLC11A1 polymorphisms and TB were identified, and full-text article retrieve excluded 11 of them  might be a risk factor for TB with a summarized OR of 1.35  as compared to TGTG+/+ genotype. Medium heterogeneity between studies  was observed. No evident publication bias was found . In the stratified analysis, the strength of the association was most evident for Asians, and marginal results were observed for Africans and Westerns. No pronounced difference was found according to different study base.2 = 61.26%) and A allele carriage was significantly associated with TB . No substantial publication bias was observed . In the subgroup analyses, significant relations were observed for Africans and Westerns.As shown in 2 = 32.47%) was observed. No significant publication bias was observed . The association was not significant any more in Westerns in the subgroup analysis.As shown in SLC11A1 5′ promoter (GT)n polymorphism was shown in 2 = 61.78%). No publication bias was found . In the stratified analyses, similar results were observed between subgroups according to study base, and significant association was found only for Africans.Meta-analysis of the association between TB and SLC11A1 polymorphisms and TB were not substantially changed n) were identified, and their effects were summarized by means of meta-analysis. Significant associations with TB susceptibility were observed for all these four loci. Strength of the associations in the subgroup analyses with respect to study population was not consistent. However, significant association has been observed for all populations  with at least one of the four loci. Limited number of studies in the stratified analysis and heterogeneity between studies might explain, at least in part, the existing inconsistency.This review addressed the associations between SLC11A1 polymorphisms and these infections and diseases has provided strong evidence for this hypothesis.TB is a serious public health problem in worldwide. Early diagnosis, drug resistance, vaccine and HIV co-infection are major factors influencing efficiency of TB prevention and control SLC11A1 polymorphisms and TB which based on 14 eligible studies published until December 2004 SLC11A1 polymorphisms. Three ethnic groups were defined with respect to the distribution of study population in the included studies, e.g. Asians, Africans and Westerns (Europeans and Americans). Statistically significant or marginal associations with TB susceptibility were found for the four studied loci in all of the three ethnic groups. Large number of included articles in the present study makes the evidence stronger to propose a consistent effect of SLC11A1 polymorphisms in different populations. In addition, population-based study has shown to be more powerful for such genetic association analysis. However, only minority of included studies (7 of 36) were conducted as population-based and no substantial different or more pronounced effect was observed as compared to those hospital-based studies.In 2006, Li HT and colleagues reported a meta-analysis on The stratified analyses suggested different study population and study base might partly explain the moderate heterogeneity between studies observed in our analyses. Further sensitivity analyses were performed as well to explore potential origin of the heterogeneity see . When reThere are some limitations to this systematic review that should be kept in mind. First, the potential confounding effect of age, sex and ethnicity was not controlled for in more than a half of included studies, which makes the interpretation of the results and stratified analyses difficult. Second, because not all necessary information could be obtained from all included studies, more detailed sub-grouping analysis (e.g. by HIV status or type of TB) could not be performed. Third, the crude division of ethnics groups into ‘Asian’, ‘African’, and ‘Western’ makes the analyses be prone to bias. Further studies from different populations are warranted to verify current findings. Fourth, included studies were restricted to those published in English or Chinese in our study which might introduce potential bias into data analysis as well. Fifth, only 17 included studies mentioned whether their study population was in HWE for the investigated variants SLC11A1 polymorphisms and TB susceptibility. Our results suggested a consistent association between SLC11A1 polymorphisms and TB in different populations, which supports the hypothesis that NRAMP1 might play an important role in the host defense to the development of TB. However, due to the moderate strength of the associations, their values to be used for risk prediction should be considered cautiously.In conclusion, this systematic review summarized the associations between SLC11A1 polymorphisms and TB were identified by searching for articles in the PubMed, EMBASE and Chinese BioMedical Literature (CBM) Database until 15 September 2010 SLC11A1”, “polymorphism” and “susceptibility” were used to screen for potentially relevant studies. Additional studies were also identified by means of cross-referencing.Studies addressing the association between SLC11A1 polymorphisms and TB were included. Only the most widely studied polymorphisms were considered: 3′ UTR (1729+55del4) (rs17235416), D543N (Asp543Asn) (rs17235409), INT4 (469+14G/C) (rs3731865), and 5′ promoter (GT)n (rs34448891). Exclusion criteria were: 1) review articles; 2) studies in languages other than English or Chinese; 2) studies on other polymorphisms other than the target four polymorphisms; 3) previous articles addressing meta-analysis of the associations between SLC11A1 polymorphisms and TB. If the eligible study was reported in duplication, the article published in English or published earlier was included in this review.Case-control or cohort studies presenting original data on the associations between For all studies, we extracted the following data from original publications: first author and year of publication; distribution of genotypes for each polymorphism among cases and controls; characteristics of the study design and the study population .2 test. Stratified analyses were conducted with respect to study base  and host ethnicity. The latter was categorized into Africans, Asians and Westerns (Europeans and Americans). Because of the limited number of publications in Americans, they were sub-grouped to Westerns combined with studies from Europeans. Begg rank correlation method and Egger weighted regression method were used to statistically assess publication bias . Sensitivity analyses were performed after excluding studies specifically on extra-pulmonary TB, or restricting the analysis specifically on pulmonary TB.Hardy-Weinberg Equilibrium (HWE) was examined in controls by asymptotic Pearson's chi-square test for each polymorphism in each study. The association between polymorphism and TB was estimated by means of odds ratios (OR) and corresponding 95% confidence intervals (CI) comparing cases to controls. Co-dominant model was used for 3′ UTR, D543N, and INT4. The effect of allele 3 carriage was assessed for 5′ (GT)n. Meta-analyses were carried out using Comprehensive Meta-Analysis . Random effects models were used for meta-analysis, taking into account the possibility of heterogeneity between studies which was tested by the Q test and ITable S1List of excluded studies and corresponding exclusion criteria.(DOC)Click here for additional data file.Table S2Characteristics of the included studies (Part 1/2).(DOC)Click here for additional data file.Table S3Characteristics of the included studies (Part 2/2).(DOC)Click here for additional data file.Table S4Sensitivity analyses of meta-analyses.(DOC)Click here for additional data file.


MTHFR polymorphism and cervical cancer risk, a meta-analysis was performed.To assess the association between MTHFR polymorphism and cervical cancer risk. The pooled odds ratio (OR) with 95% confidence intervals (CIs) were calculated.Based on comprehensive searches of the PubMed, Embase, and Web of Science databases, we identified outcome data from all articles estimating the association between MTHFR C677T polymorphism and CIN I risk . For the CIN II/III, lack of an association was also found . The T allele had significant association to susceptibility of invasive cervical cancer in recessive model . On subgroup analysis by ethnicity, similarly significant differences in T vs. C, TT vs. CC, and recessive model were found in Asians.A total of 12 studies with 2,924 cases  I, 742 CIN II/III, 1851 invasive cervical cancer) and 2,581 controls were identified. There was no significant association between MTHFR C677T polymorphism were to substantially contribute to invasive cervical cancer in recessive model.The present meta-analysis suggested that  Cervical cancer continues a serious threat to women throughout the world MTHFR gene might be one of the candidate genes for susceptibility of cervical cancer.Methylenetetrahydrofolate reductase (MTHFR), a homodimeric enzyme, catalyzes the conversion of 5,10-methylenetetrahydrofolate to 5-methyltetrahyd- rofolate MTHFR C677T polymorphism and cervical cancer risk. However, the MTHFR C677T polymorphism’s association with cervical cancer, or the lack thereof, remain inconclusive. To derive a more comprehensive and precise estimation of the relationship, we carried out a meta-analysis on all eligible case-control studies to estimate the effect of MTHFR polymorphism on the risk of cervical cancer.A relatively large number of studies evaluated the association between MTHFR C677T polymorphism and susceptibility of CIN I MTHFR C677T polymorphism and susceptibility of CIN II/III MTHFR C677T polymorphism and susceptibility of cervical cancer Twelve publications, including 2,924 cases  I patients, 742 CIN II/III patients, 1851 invasive cervical cancer patients) and 2,581 controls, met the inclusion criteria Fixed effects models were used to calculate the pooled OR in all genetic models. Overall, the combined results showed that no significant association was found in all genetic models . Forest plots on the basis of all studies were shown in MTHFR C677T polymorphism indicated that the T allele had no significant association to CIN II/III susceptibility as compared to the C allele under the fixed effects models , TT vs. CC, , and TT+CT vs. CC . The combined results showed significant differences in recessive model . When stratified by ethnicity, we observed a wide variation of T allele frequencies between the controls across different ethnicities. The result of One-way ANOVA indicated that the T allele frequencies were significant difference in Caucasians, Asians, and Mixed populations (P = 0.015). When meta-analysis was performed to assess association between MTHFR C677T polymorphism and different ethnicities, the T allele of MTHFR C677T polymorphism had significant association with invasive cervical cancer susceptibility in Asians. The results were showed in MTHFR C677T polymorphism and invasive cervical cancer susceptibility, there were statistically significant heterogeneity in T vs. C , TT vs. CC , dominant genetic model , and recessive genetic model .For the association between I2 values were less than 50% and PQ were greater than 0.1 . Two studies with relatively low NOS score (<7) were excluded from the sensitivity analysis. The sensitivity analysis indicated the results of our meta-analysis were relatively consistent even when some studies were excluded. The results were shown in Publication bias was estimated by the funnel plots. As shown in MTHFR in maintaining folate homeostasis, the MTHFR C677T polymorphism has been investigated in certain types of cancer, which included Colorectal, Thyroid, Breast, Ovarian, and cervical cancers MTHFR C677T polymorphism and cervical cancer risk was first reported in a mixed populations by Piyathilake et al MTHFR in cervical cancer susceptibility and presentation have been reported by series of case-control studies MTHFR C677T polymorphism and cervical cancer risk.Worldwide study has indicated that folate levels show a protective role in a variety of cancers. Owing to the importance of MTHFR C677T polymorphism were performed to provide the most comprehensive assessment of the relationship between polymorphism and cervical cancer risk. The T allele of MTHFR C677T polymorphism had no association with the CIN I susceptibility for the T vs. C, TT vs. CC, dominant genetic model, and recessive genetic model in overall populations. Lack of an association was also found in CIN II/III and cervical cancer. In view of the complex effect of genetic polymorphisms on disease progression, the lack of an association between MTHFR C677T polymorphism and invasive cervical cancer susceptibility may attribute to other polymorphisms in MTHFR gene promoter which could affect the activity of MTHFR. Ulvik A et al. MTHFR A1298T polymorphism was associated with reduced MTHFR activity. Meanwhile, the MTHFR C677T and A1298T polymorphisms appeared to interact with folate in determining cancer risk. Strong correlation between MTHFR C677T and A1298T polymorphisms was observed in cervical dysplasia as compared to normal cervical cytology MTHFR A1298T polymorphism and cervical cancer risk. There was no association between MTHFR A1298T polymorphism and cervical cancer risk  on cer risk  and S2. MTHFR C677T polymorphism and invasive cervical cancer susceptibility, subgroup analysis by ethnicity was performed. First, we detected whether there was T allele frequency of variation in different ethnicities. The T allele frequency has significant differences in different populations. Next, the association between MTHFR C677T polymorphism and invasive cervical cancer risk in different ethnicities was explored. Lack of an association was also found in all genetic models.To explore a more precise relationship between MTHFR C677T polymorphism and invasive cervical cancer risk. Then, we used the Galbraith plots to explore the sources of heterogeneity. We found all of the I2 values were less than 50% and PQ were greater than 0.1 after excluding the studies of Shekari M et al., Ma XC et al., and Zoodsma M et al. respectively. The results indicated that the three studies might be the major source of the heterogeneity for the association between MTHFR C677T polymorphism and cervical cancer risk. The results of subgroup analysis revealed that the ethnicity might contribute to the potential heterogeneity.In our meta-analysis, obvious heterogeneity was observed for the association between There are some limitations to this meta-analysis. Firstly, the retrieved literature is potentially not comprehensive enough. Studies included in our meta-analysis were limited to published articles. We did not track the unpublished articles to obtain data for analysis. Secondly, as many other factors such as age, parity, smoking, and alcohol consumption may participate in the progression of disease, we did not carry out subgroup analysis based on these factors due to limited data. Thirdly, the small sample sizes in some subgroup analyses limited the ability to draw more solid conclusions.MTHFR C677T polymorphism may associate with genetic susceptibility of invasive cervical cancer in recessive model based on the current published studies. Similarly significant differences in T vs. C, TT vs. CC, and recessive model were found in Asians. Moreover, further studies with large sample size of different ethnic populations will be necessary to combine genetic factors together with age, parity, smoking, and alcohol consumption.Conclusively, This meta-analysis followed the Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA) criteria MTHFR C677T polymorphism and cervical cancer or its precursor lesion, CIN, (2) case-control studied, (2) sufficient data for estimating an odds ratio (OR) with 95% confidence intervals (CIs), (3) genotype distribution of control population in HWE, and (4) studies written in English or Chinese. For the studies with the same or overlapping data by the same authors, the most recent or largest population was selected.Two reviewers independently screened titles and abstracts of all studies for relevancy. Disagreements were resolved by a third opinion. Full-text publications were retrieved for relevant articles. The strength of the individual studies was weighed for relevance, based on the following items: (1) evaluation of the Data were extracted independently from each study by two reviewers according to the inclusion criteria listed above. Agreement was reached after discussion for conflicting data. The following data were collected from each study: first author’s name, publication year, original country, ethnicity, control source, sample size, genotyping method, and genotype number in cases and controls.The quality of included studies was assessed independently by the same two investigators using the NOS MTHFR polymorphism and susceptibility of cervical cancer or CIN was estimated by OR and corresponding 95% CIs. The pooled OR was calculated respectively for T vs. C, TT vs. CC, dominant genetic model (TT+CT vs. CC), and recessive genetic model (TT vs. CT+CC). Between-study heterogeneity was assessed by the Q-test and I2 test, PQ <0.10 and I2>50% indicated evidence of heterogeneity. Then, the random-effects model (the DerSimonian and Laird method)The strength of association between Subgroup analyses were performed by ethnicity of study population. Sensitivity analysis was performed based on the high quality studies (according to the NOS score). Asymmetry funnel plots were inspected to assess potential publication bias. The Egger’s linear regression test was also used to assess publication bias statistically Data were analyzed by using STATA 11.0  and Revman 5.0 (The Cochrane Collaboration).Figure S1Flow diagram of the selection of eligible studies.(TIF)Click here for additional data file.Figure S2Funnel plots of all genetic models in overall studies. A. T vs. C; B. TT vs. CC; C. dominant model (TT+CT vs. CC); D. recessive model (TT vs. CT+CC). Funnel plots of dominant model seemed asymmetry. Each point represents a separate study for the indicated association.(TIF)Click here for additional data file.Table S1Association between individual study characteristics and MTHFR A1298T polymorphism.(DOC)Click here for additional data file.Table S2Meta-analyses of MTHFR A1298T polymorphism and risk of cervical cancer.(DOC)Click here for additional data file.PRISMA S1Checklist Association between methylenetetrahydrofolate reductase C677T polymorphism and susceptibility of cervical cancer.(DOC)Click here for additional data file.


A state of low-grade inflammation accompanies the pathogenesis of atherosclerotic events. Interleukin-6 (IL-6) is a pleotropic pro-inflammatory cytokine that modulates the development of acute coronary syndromes (ACSs), partly by destabilizing coronary atherosclerotic plaques. We have examined the contribution of the -174G>C IL-6 promoter variant on the risk of coronary artery disease (CAD) among Tunisians.Study subjects included 418 CAD patients and 406 age- and sex-matched controls. IL-6 genotyping was done by PCR–restriction fragment length polymorphism.The frequency of the -174C allele (mutant) was lower in Tunisians than in Europeans, and the distribution of -174 G>C genotypes was similar between CAD patients and control subjects. Moreover, compared to GG genotype carriers, -174C allele carriage did not increase the CAD relative risk , which remained nonsignificant after adjusting for traditional risk factors for CAD .The -174G>C IL-6 promoter variant is not associated with an increased risk of CAD among Tunisians. Coronary artery disease (CAD) is a major cause of death and disability in developed countries, and is characterized by a long asymptomatic developmental phase, which progresses to precipitation of atherosclerotic plaques, the fatty streak being the earliest visible lesion.2Interleukin-6 (IL-6) is a 26-kDa pleotropic inflammatory cytokine produced by many cell types, including fibroblasts, monocytes, adipocytes and endothelial cells558131315Polymorphic variants in the IL-6 gene were implicated in the expression of acute-phase proteins and were described as regulators of atherogenic markers. Of these, the commonly studied polymorphism is the functional variant -174G>C. Association of this single nucleotide polymorphism (SNP) with CAD has been reported in some but not all the studies.2 or higher. Hypertension was defined as blood pressure  >140/90 mm Hg on two separate occasions, measured using mercury sphygmomanometer with participants in the sitting position following a 5-minute period of rest (the mean of two readings measured 1 minute apart was adopted) and/ or the use of antihypertensive therapy . Diabetes was diagnosed according to WHO criteria (fasting blood glucose >12.6 mg/dL or 70 mmol/L) on at least two occasions and/or the use of glucose-lowering drugs, including insulin).This was a case-control study involving 418 CAD patients  diagnosed as having CAD based on criteria given by the World Health Organization (WHO), and who attended the Farhat Hached Hospital in Sousse, Tunisia. Diagnosis of CAD was based on coronary angiography . Determination of CAD severity was based on the number of major epicardial coronary arteries affected. Exclusion criteria included concomitant renal, hepatic or autoimmune disease. Obesity was defined as body mass index (BMI) of 30 kg/mThe control population was comprised of 406 healthy subjects  who either attended a routine health check at a general practice or at their workplace. They were required to have a normal physical examination, a normal resting electrocardiogram and no personal or family history of CAD. All participants were asked to fill a standard questionnaire detailing demographic details, together with data on traditional CAD risk factors. All subjects were asked to sign a written consent form after the aims and details of the study were explained to them. The study was approved by the Ethics Committees of the University Hospital of Sousse, Tunisia.Blood for IL-6 genotyping was collected in EDTA tubes and kept frozen pending DNA extraction. Total genomic DNA was extracted from the buffy coat layer by the salting-out procedure and was stored in TE buffer . Genotyping of -174G>C (Nla III) was performed by PCR-restriction fragment length polymorphism (PCR-RFLP) as described.t test was used for analyzing continuous variables , and chi-square analysis was employed for analyzing categorical variables ; a P value <.05 was considered statistically significant. The associations of specific genotypes with CAD were determined by logistic regression analysis, adjusted for age, smoking, hypertension, diabetes and body mass index (BMI). Taking homozygous carriers of the wild-type allele as reference, two independent odds ratios (ORs) were estimated, one for heterozygote carriers and the other for homozygote carriers.Statistical analysis was carried out using SPSS version 13.0 software . Data were expressed as mean (standard deviation) for continuous variables or as percentages of the total for categorical variables. The Table 1 lists the demographic and clinical characteristics of the 418 CAD patients and 406 control subjects. The two groups were matched for gender (P=.06) and age (P=.11). CAD patients had a higher BMI (P<.001), systolic (P<.001) and diastolic (P=.006) blood pressure, and higher total cholesterol (P<.001), triglyceride (P<.001), creatinine (P<.001) and urea (P<.001) levels and lower HDL-cholesterol levels (P<.001). The prevalence of hypertension (P<.001) and rate of smoking (P<.001) were higher among patients than among control subjects.Table 2). No statistically significant differences in genotypes or allele distribution were seen between patients and controls, and -174 G>C did not exhibit transmission distortion. The distribution of the minor allele -174C (15.5% vs. 14.3%), and -174G/C (26.3% vs. 25.1%) and 174C/C (2.4% vs. 1.7%) genotypes was comparable between patients and controls, respectively. An increased prevalence of -174C allele was seen among patients with unstable angina (16.1%) or myocardial infarction (17.0%) patients compared to stable angina (8.6%) patients (Table 3).Genotype distribution of the -174G>C polymorphism did not deviate from the Hardy-Weinberg equilibrium among participants (Table 4). This lack of association was irrespective of whether a dominant or recessive effect of the allele was assumed.The association of -174G>C IL-6 variant with CAD was examined first by univariate regression analysis and later by multivariate regression analysis. Taking -174G/G genotype as the reference, the univariate regression analysis demonstrated no significant association of -174G/C genotype  or -174C/C genotype  with CAD. Multivariate regression analysis confirmed the lack of association of -174G/C genotype  or -174C/C genotype  with CAD, after adjusting for CAD traditional risk factors   and -174C/C (2.4) genotypes in our patient group was lower than that in French or Irish (Belfast) population, as reported in the Etude Cas Temoin de l’Infarctus du Myocarde (ECTIM) study,19.3 and -128*C allele in patients with myocardial infarction (compared with patients with stable and unstable angina), suggesting that the -174G/C variant may be involved in CAD pathogenesis.29Regression analysis confirmed the lack of association of -174G/C genotypes with CAD. Our results are in accord with an earlier study demonstrating the lack of association of the G/C genotype, and a weak but statistically nonsignificant association of the C/C genotype, with increased CAD riskGiven the well-established role of inflammatory mechanisms in CAD pathogenesis, coupled with the role of IL-6 as a pro-inflammatory cytokine, it is tempting to speculate that induction of inflammation mediated by altered IL-6 levels regulated by specific IL-6 variants may modulate CAD development and progression. Taken together, these findings suggest that the carriage of the -174C allele imparts a weak CAD association, compared to the -174G/G genotype carriers. Our findings suggest that genetic variations in the IL-6 gene are unlikely to play an important role in the genetic predisposition to CAD, and that it represents a poor risk indicator in patients with CAD.The strengths of this study lie in the sample size used (418 patients and 406 control subjects), which was sufficiently powered to reduce type I errors; and in being the first to examine the association of IL-6 promoter variants with CAD in the ethnically homogenous Tunisian population. However, our study has some limitations; namely, that we were not able to measure plasma IL-6 concentrations in patients and controls, and that it was limited to a specific ethnic group (north African Tunisian Arabs), thereby necessitating follow-up studies in CAD patients from different ethnic groups. Another limitation of the study is that IL-6 genotyping was determined in confirmed CAD patients, thus questioning the value of analyzing these variants, as it is not clear whether mutation-carrying control subjects may develop future CAD events. The possible involvement of other IL-6 variants, as well as nearby or distant functional gene polymorphisms, in disease pathogenesis remains to be seen. Our study is of potential interest since comparison of genetic associations among different populations with possibly different SNP frequencies is useful.