LTA gene and susceptibility to MI have produced contradictory and inconclusive results. The aim of this meta-analysis is to provide a relatively comprehensive account of the association of these polymorphisms with susceptibility to MI.The lymphotoxin-α (LTA), as one of the mediators of inflammation, may play an important role in the pathogenesis of myocardial infarction (MI). Genetic association studies (GAS) that have investigated the association between three common polymorphisms  of the A literature search for eligible GAS published before October 15, 2013 was conducted in the PubMed, Embase, Web of Science, Cochrane Library, and CNKI  databases. We performed a meta-analysis of fifteen case-control studies with a total of 22,549 MI patients and 16,105 healthy controls.LTA A252G, a borderline significant overall association was found, indicating that GG genotype may confer an increased susceptibility to MI compared to AA and AG genotypes. Based on an ethnicity stratification analysis, a significant association was observed in Asians, but not in Caucasians. For LTA G10A, no significant overall association was found. However, subgroup analysis based on ethnicity suggested that the 10A allele may confer a significant increased susceptibility to MI only in Asian populations. For LTA C804A, the combined results revealed a significantly increased susceptibility to MI for carriers of the 804A allele in both overall analysis and stratified analyses.For LTA C804A may be associated with an increased susceptibility to MI, whereas LTA A252G and G10A may confer a significant increased susceptibility to MI only in Asians. Thus, these polymorphisms of the LTA gene can probably be used with other genetic markers together to identify individuals at high susceptibility to MI especially in Asians.This meta-analysis shows that  Myocardial infarction (MI) remains the principal cause of death in many countries despite improvements in lifestyle and the development of new pharmacologic approaches Lymphotoxin-α (LTA) is one of the cytokines produced in the early stages of vascular inflammatory processes LTA gene, which encodes LTA on chromosome 6p21, has been linked with the risk of MI LTA gene seem to be involved in inflammation by both qualitatively and quantitatively modifying the function of the LTA protein LTA gene for genetic association with MI risk, including A252G (dbSNP: rs909253) in intron 1, G10A (dbSNP: rs1800683) in exon 1 and C804A (dbSNP: rs1041981) in exon 3. Their results, however, have proven conflicting. In an initial genome-wide case-control screen involving 65,671 single nucleotide polymorphisms (SNPs) from 13,738 genes in 1,133 MI cases and 1,006 controls in Japan, susceptibility to MI appeared to be associated with the LTA A252G polymorphism LTA gene in relation to MI, we performed a meta-analysis using published data from observational studies.The LTA genetic polymorphisms and susceptibility to MI published before October 15, 2013 was conducted in the following electronic databases: PubMed, Embase, Web of Science, Cochrane Library, and CNKI  databases. The following combinations of main keywords were used:  and  and (‘genetic polymorphism’ or ‘single nucleotide polymorphisms’ or ‘SNP’). The search was done without limitations on language but only included those studies that were conducted on human subjects. All references in eligible articles were extensively reviewed to identify additional published articles.A literature search for GAS that investigated the association between the LTA genetic polymorphisms and susceptibility to MI; (2) all patients in the candidate studies meet the diagnostic criteria for MI; (3) studies with sufficient available data to calculate ORs with corresponding 95%CIs. The major reasons for excluding studies were: (1) not case-control study; (2) duplicate publications with overlapping subjects from the same study; and (3) no available data reported. For multiple studies using overlapping cases or controls, the most recent study with the largest sample size was included in the meta-analysis. This meta-analysis was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidance with only slight modification, and did not require ethics board approval  case-control studies on the association between the approval  [24].According to the PRISMA guidance, two investigators independently checked each full-text report for eligibility and extracted the following data from eligible studies: surname of first author, year of publication, country of origin, ethnicity, definition and number of case and control, age, sex ratio, genotyping method, allele and genotype frequency, etc. Disagreements were solved by discussion between all authors until consensus was reached. For data not provided in table form or in the main text, required information was obtained by contacting corresponding authors when possible.The strengthening report of genetic association studies (STREGA) quality score system and the Newcastle-Ottawa Scale (NOS) criteria were used to assess the qualities of all included studies LTA genetic polymorphisms and susceptibility to MI was assessed by the pooled odds ratios (ORs) with their corresponding 95% confidence intervals (95%CIs) under five genetic models, including the allele model, the dominant model, the recessive model, the homozygous model and the heterozygous model. Taking into consideration possible between-study heterogeneity, a statistical test for heterogeneity was first conducted using Cochran's Q statistic and the 2I metric 2I exceeding 50% as an indicator of significant heterogeneity. When no heterogeneity was found with P>0.10 or 2I<50%, a fixed-effects model was used to estimate the pooled ORs and 95%CIs. Otherwise, a random-effects model was applied. In addition to an overall comparison, stratified analyses, based on ethnicity, source of control, HWE status, and genotyping method where applicable, were also performed to explore possible explanations of between-study heterogeneity and to investigate whether overall reported associations were present in subgroups. Univariate and multivariate meta-analyses were conducted to explore potential sources of heterogeneity, considering publication year, ethnicity, genotyping method, source of control, and quality score. Sensitivity analyses were conducted by omitting individual studies in turn to reflect the influence of individual datasets on the pooled results P<0.05 were considered statistically significant, and all analyses were performed using the STATA 12.0 software .Genotype distributions in the controls were tested for conformation to Hardy-Weinberg equilibrium (HWE) using the chi-square test. HWE in the controls was tested by comparing the expected and observed genotype frequencies using the Pearson chi-square test for goodness of fit. The association between the LTA A252G polymorphism and susceptibility to MI is summarized in P<0.10 and 2I>50% under all genetic models), the random-effects model was used. As shown in P = 0.040; GG vs. AA: OR = 1.21, 95%CI: 1.01–1.45, P = 0.039). However, exclusion of one hospital-based study by Yamada et al. made the summary ORs in population-based studies become insignificant, and after excluding of one non-HWE study by Ozaki et al., the pooled ORs in HWE studies also became insignificant, suggesting that the A252G polymorphism is unlikely to have a major role in the risk of MI. Interestingly, in a stratified analysis by ethnicity, we found that this polymorphism played different roles in Asian and Caucasian populations. In Asians, subjects harboring the 252G variant are approximately 20% more likely to have a MI when compared to subjects with the 252A allele . However, no significant effects were found for the Caucasians  , whereas no significant association was found in the non-PCR-RFLP subgroup . Univariate and multivariate meta-analyses further confirmed the above results of subgroup analysis, which suggested that ethnicity might be the major source of between-study heterogeneity with 87% was explained by this covariate  . Stratifovariate .LTA G10A polymorphism and susceptibility to MI is provided in P<0.10 and 2I>50% under all genetic models). The overall analysis showed that LTA G10A may not be associated with the risk of MI under any of the five genetic models . Exclusion of one non-HWE study by Ozaki et al. did not make the summary ORs significant in HWE studies. However, stratified analyses based on ethnicity suggested that the 10A allele may confer a significantly increased susceptibility to MI in Asians , but not in Caucasians, as shown in A summary of findings on the relationship between the LTA C804A polymorphism and susceptibility to MI. Nine case-control studies had investigated the relationship between C804A and susceptibility to MI with a total of 19,404 MI patients and 13,684 healthy controls. Since between-study heterogeneity obviously existed , the random-effects model was used. Overall, the LTA C804A polymorphism was found to be associated with a significant increased susceptibility to MI . As shown in LTA gene involved in the current study, stratified analysis based on ethnicity revealed a significant association in Asians, but not in Caucasians , whereas no significant association was found in the other subgroups . Similar to A252G, univariate and multivariate meta-analyses also suggested that ethnicity might be the major source of between-study heterogeneity for C804A with 83% was explained by this covariate . Begg's funnel plots and Egger's linear regression test were used to assess the potential publication bias of included studies under the allele model. The shapes of the funnel plots did not reveal any evidence of obvious asymmetry . The above tests indicated a promising level of robustness and accuracy for the results of this meta-analysis.Sensitivity analyses for symmetry . In addiLTA gene and the LTA protein are associated with MI onset. LTA is a proinflammatory cytokine with homology to inflammatory cytokine tumor necrosis factor alpha (TNF-α) and is related with the development of atherosclerotic lesions in coronary arteries LTA gene can modify the function of the LTA protein qualitatively and quantitatively, thereby conferring a risk for MI, as shown in LTA gene, such as A-to-C in intron 1, G-to-A in exon 1, and C-to-A in exon 3, may influence the inflammatory biological activities through causing functional mutations in LTA protein, which in turn affect the degree of inflammation and confer a higher susceptibility to MI than normal LTA protein. Based on this fact, several investigations have been conducted to assess the association between common LTA genetic polymorphisms and susceptibility to MI. Previously, Padovani et al. first investigated the association between LTA A252G and susceptibility to MI in a Brazilian population, but failed to find a significant association LTA genetic polymorphisms on susceptibility to MI in Japanese populations LTA A252G and C804A on susceptibility to MI in a Taiwanese population LTA A252G, G10A and C804A polymorphisms and susceptibility to MI.MI is a multifactorial disease significantly associated with certain genetic factors LTA gene and various diseases, including gastric cancer LTA genetic polymorphisms with susceptibility to MI. This systematic review provides a more comprehensive summary of the currently available evidence on the associations between the LTA A252G, G10A and C804A polymorphisms and susceptibility to MI. In this meta-analysis, the LTA C804A polymorphisms seem to be associated with an increased susceptibility to MI, whereas LTA A252G and G10A may confer a significant increased susceptibility to MI only in Asians.Meta-analysis has the advantage of synthesizing data from published GAS to obtain greater statistical power for detecting significant associations than available in an individual GAS, especially in the absence of large heterogeneity between studies LTA genetic polymorphisms played different roles in Asian and Caucasian populations. All these LTA genetic polymorphism had significant increased effects on the risk of MI in Asians, whereas no significant effects were found in Caucasians. These conflicting results may be due to the different genetic backgrounds of these populations, subsequently leading to different genetic susceptibility to the same disease. In addition, the source of controls was another factor that contributed to heterogeneity. The genotype distribution in population-based controls may be similar to normal and thus population-based controls could be more reliable than hospital-based controls. This might partially explain why the results of the stratified analysis by the source of the controls showed differences between the two subgroups for LTA A252G. Thus, further studies with larger sample size using population-based controls are warranted. Moreover, subgroup analysis based on genotyping method showed a significant association between LTA A252G and C804A and susceptibility to MI in the PCR-RFLP subgroup, whereas no significant association was found in the non-PCR-RFLP subgroups. This result may be due to the overlapping effect of ethnicity, since the significant association was also observed in Asians and most included studies on Asian subjects used the PCR-RFLP genotyping method. In addition, this association may also be due to chance especially for C804A, for which only three studies were involved in the PCR-RFLP subgroups. Hence, further investigations are also warranted to assess the effect of different genotyping methods on the significance of the association between LTA genetic polymorphisms and susceptibility to MI.Heterogeneity is a major problem when interpreting the results of meta-analyses. As the results of meta-regression shows, ethnicity was an important source of this heterogeneity as individuals of different ethnicities may have diverse genetic backgrounds and environmental factors, and as a result the same polymorphism may play different roles in different populations. Interestingly, our subgroup analysis by ethnicity showed that the LTA A252G and G10A polymorphisms should be interpreted in caution. For A252G, exclusion of hospital-based and non-HWE studies made the summary ORs become insignificant, suggesting that this polymorphism is unlikely to have a major role in susceptibility to MI. For G10A, the significant association in Asians should also be treated carefully, since only two studies on Asians were involved and thus statistical power was limited due to the small sample size. Hence, the significant associations of LTA A252G and G10A with susceptibility to MI should be interpreted in caution and more large-scale studies are warranted for confirming these relationships. Second, as with other complex traits, susceptibility to MI may be modulated by other genetic markers besides the LTA gene. Thus, fully elucidating the pathogenesis of MI would demand an investigation into the association and combined interaction of many gene variants with susceptibility to MI. Third, the included studies only focused on the Asian and Caucasian populations and thus further studies on a wider spectrum of subjects should be carried out to investigate the role of these variants in different ethnicities. Last, this meta-analysis was based on unadjusted ORs and possible effect modifiers, such as sex, age, BMI, diabetes mellitus, and smoking status, may influence the estimates of associations. Unfortunately, the calculation of adjusted pooled ORs and further subgroup analyses based on these factors could not be performed because of limited data. Thus, further well-designed GAS need to focus on exploring these sources of heterogeneity. Despite these limitations, our study is the first comprehensive meta-analysis of all eligible studies on the association between the LTA genetic polymorphisms and MI risk.In interpreting the results of this meta-analysis, some specific issues need to be addressed. First, the significant associations found for the LTA A252G and C804A polymorphisms may be associated with an increased risk of MI, whereas LTA G10A may confer a significant increased risk of MI only in Asian populations. Thus, these polymorphisms of the LTA gene could possibly be used with other genetic markers together to identify individuals at high risk for MI. However, due to the limitations of this study, these results should be interpreted with caution and still require future large-scale studies to confirm their accuracy. Moreover, considering that MI is a complex disease with a multifactorial etiology, the development of MI might be associated with gene-gene and gene-environment interactions, whose effects should be considered in future GAS and subsequent meta-analyses that may provide more conclusive evidence regarding the genetic susceptibility to MI.In summary, the current meta-analysis indicates that the Supplement S1The Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) Checklist.(DOC)Click here for additional data file.Supplement S2The Strengthening the Reporting of Genetic Association Studies (STREGA) quality score systems.(DOC)Click here for additional data file.Supplement S3The Newcastle-Ottawa Scale (NOS) criteria.(DOC)Click here for additional data file.Supplement S4Univariate and multivariate meta-analyses of potential source of heterogeneity.(DOC)Click here for additional data file.


TLR9) and susceptibility to cancer, however, the results remained inconsistent.Growing studies have revealed the association between polymorphisms in the Toll-like receptor 9  and corresponding 95% confidence intervals (CIs) for polymorphisms in TLR9 and cancer risk were estimated. Our meta-analysis indicated that rs352140 was associated with an increased cancer risk, especially in Caucasian. However, no significantly increased cancer risk was detected to be associated with rs187084 and rs5743836 either the overall or subgroup estimation.To assess the effect of three selected SNPs  in TLR9 may play a role in cancer development.These meta-analysis results indicate that polymorphisms in  Toll-like receptors (TLRs), expressed predominantly on antigen-presenting cells, belongs to the family of pattern-recognition receptors (PRRs). In humans, TLR family consists of 10 members (TLRs 1–10) TLR9 is located on chromosome 3p21.3 TLR9 gene polymorphisms in various cancers, including bladder cancer Human TLR9 with cancer risk. Thus, we performed this meta-analysis with accumulated data to evaluate the overall cancer risk of selected three SNPs in TLR9 and to quantify heterogeneity between the individual studies as well as to investigate the existence of potential publication bias.Considering a single study might underpowered to detect the overall effects in complex diseases, a quantitative synthesis of the accumulated data from different studies was deemed important to provide evidence on the association of variants in TLR9 polymorphisms and Cancer risk  using the following terms: ‘Toll like receptor 9’ or ‘TLR9’ and ‘cancer’ or ‘tumor’ or ‘carcinoma’ and ‘polymorphism’ or ‘polymorphisms’ or ‘SNP’ for relevant reports. In order to identify the relevant publications, the references cited in the research papers were also scanned.We searched PubMed and CNKI  for all articles on the association between TLR9 polymorphism and cancer risk, (2) being a case-control study, (3) existing useful genotype frequency , (4) control subjects satisfied the Hardy-Weinberg equilibrium (HWE), and (5) the study was published in English or Chinese. Abstracts and unpublished reports were not considered.The inclusion criteria for current meta-analysis studies were (1) evaluation of the Data extraction was independently done by two investigators (Zhang L.S. and Qin H.J.), and discrepancies were resolved by consensus including a third investigator (Zhang K.). The following characteristics were collected from each study: first author, year of publication, country of the study population, ethnicity, cancer types, genotyping methods, number of genotypes, sample size, minor allele frequency (MAF) in controls, and evidence of Hardy-Weinberg equilibrium (HWE). Eligible studies were stratified into population-based (PB) and hospital-based (HB) according to the control source. Once studies include subjects of different ethnic groups, data were extracted separately for each ethnic group.The quality of eligible studies was evaluated by three reviewers  independently by scoring according to a ‘methodological quality assessment scale’ , which wP<0.05 was considered as significant disequilibrium. Strength of association between TLR9 polymorphism and cancer risk was evaluated by crude odds ratio (OR) and 95% confidence interval (CI). Pairwise group differences of ORs were analyzed and the best genetic models were to be determined according to the Thakkinstian’s method HWE in controls for each study was assessed by using a goodness of fit chi-square test before statistical analysis and P>0.1, then ORs were pooled according to the fixed-effects model (the Mantel-Haenszel model) A chi-square-based Q test was used to check the statistical heterogeneity Our initial search identified 43 studies according to the search words, and 2 records added through the reference scan. After screening the abstract, 24 were retrieved for more detailed evaluation. After reviewing full text, 10 studies were excluded for the following reasons, three papers were reviews For control subjects, the MAF ranges from 0.39 to 0.66 in rs352140, from 0.08 to 0.38 in rs5743836 and from 0.21 to 0.45 in rs187084. Overall, for rs352140, rs5743836 and rs187084, no significant difference between cancers and controls were detected in allele comparison .P values were 0.178, 0.577 and 0.053, respectively)  among Caucasian, but not in Asian , OR2 (CT versus CC) and OR3 (TT versus CT) were 1.414 , 0.937  and 1.309 , and all of them were not significant (ctively) . Thus, win Asian .P values were 0.621, 0.946 and 0.971, respectively) , OR2 (CT versus CC) and OR3 (TT versus CT) were 1.054 , 0.992  and 1.008 , and all of them were not significant (ctively) . So, we ctively) .Due to the missing of the minor allele in some studies, it was difficult to get the OR1, OR2, and OR3. Hence, we performed the allele comparison and dominant comparison to evaluate the association between rs187084 polymorphism and cancer risk. However, we did not found any significant association between this SNP and the cancer risk in subgroup analysis by cancer types (lymphoma and other cancers) .2 = 54.9%, Pheterogeneity = 0.084, dominant model: I2 = 58.9%, Pheterogeneity = 0.063 for rs187084; C vs. T: I2 = 82.6%, Pheterogeneity<0.001, dominant model: I2 = 85.1%, Pheterogeneity<0.001 for rs5743836; respectively) and subgroup analysis. To explore sources of heterogeneity across studies, we assessed allele comparison by frequency of minor allele in control subjects, frequency of minor allele in case subjects, cancer types, ethnicity and sample size, when it was available. As a result, frequency of minor allele in control subjects of rs187084 could explain 82.35% of the heterogeneity. Frequency of minor allele in control subjects of rs5743836 and cancer types could explain a total proportion of 62.2% to the heterogeneity.Heterogeneity was detected among studies in overall comparisons  (Begg’s funnel plot and Egger’s test were performed to assess publication bias of the selected literature. No evidence of publication bias in our study was observed (Begg’s test ctively) .TLR9 polymorphisms and cancer risk among 14,091 subjects. Our meta-analysis identified that elevated cancer risk was statistically associated with TT genotype in recessive model of rs352140. Moreover, in terms of stratified analysis by ethnicity, allele T and genotype TT were found to increase cancer risk in both allele comparison and recessive model among Caucasian. However, no significant associations were found between rs187084 and rs5743836 polymorphisms and cancer risk either in the overall comparison or in subgroup analysis.In this study, we performed a meta-analysis to explore the association between TLR9. Previous research stated that rs352140 TT genotype was associated with a higher expression of TLR9 at the mRNA level The synonymous rs352140 polymorphism locates in exon 2 of TLR9TLR9TLR9 triggering, suggesting that the distinct cell death responses upon CpG oligodeoxynucleotides (CpG ODN) treatment in BL cells may be linked to rs5743836. Moreover, C allele of the rs5743836 polymorphism generates an IL6 responding element. In mononuclear cells carrying the CT genotype of rs5743836, IL6 up-regulates TLR9 expression, leading to exacerbate cellular responses to CpG, including IL6 production and B cell proliferation. In addition, previous study observed that C allele of rs5743836 create a putative NF-kB binding site and exhibits a greater NF-kB binding affinity which leads to enhanced transcription of NF-kB and results in greater release and production of pro-inflammatory mediators TLR9 triggering and act as an important role in host immune response, inflammation and tumorigenesis. Although some studies reveal the relationship between rs5743836 polymorphism with tumorigenesis, the evidence from epidemiological studies seems controversial. Most previous studies did not show any association between rs5743836 polymorphism and cancer risk The SNP rs5743836, substituting tyrosine to cytosine, locates in the promoter region of TLR9, created a putative Sp1 binding site, which may be functionally relevant TLR9The SNP rs187084 located in the promoter of For heterogeneity, we found the frequency of minor allele in case subjects was the main source of heterogeneity for rs187084. While, the cancer types and frequency of minor allele in control subjects were the main sources of heterogeneity for rs5743836. Thus, further studies using larger numbers of participants from multiple discrete regions and sufficient cancer types should be conducted.TLR9 polymorphisms with cancer risk and a more precise adjusted OR for other covariates like age, family history, infect condition and environmental factors were unavailable. Secondly, the studies evaluated cancer risk with rs352140 and rs187084 were limited. Hence, sub-group analysis with the specific cancer type did not performed due to insufficient data. Thirdly, for rs5743836, most participates were Caucasians. Lack of data from other ethical group may lead to an improper result. Furthermore, due to the low frequency of the minor allele, it is difficult for analysis by using any more genetic models for rs5743836. Finally, we did not found an appropriate genetic model to assess the association between rs352140 and the risk of cancer according to the Thakkinstian’s method. Thus, our results derived from genetic model of rs352140 should be debatable. In spite of these limitations, this meta-analysis with methodological quality assessment was applied and all studies included in this meta-analysis met our selection criteria clearly. Allele comparison and genetic models comparison were used to evaluate the cancer risk with TLR9 polymorphisms. Secondly, subgroups analysis by ethnicity, cancer types and sample size provided better knowledge about TLR9 polymorphisms and cancer risk.Some limitations of this meta-analysis should be monitored. First, most of the enrolled studies only included the association of TLR9 rs352140 was associated with increased cancer risk, especially in Caucasians, suggested that polymorphisms in TLR9 may play a role in cancer development. Since genetic diversities and cancer type were the main sources of the heterogeneity, it is critical that larger and well-designed multicenter studies based on other area populations and sufficient cancer types should be performed to re-evaluate the association. Moreover, additional future studies should combine with other potential risk factors to extend our investigations.In conclusion, our meta-analysis indicated that Table S1PRISMA 2009 checklist.(DOC)Click here for additional data file.Table S2Scale for methodological quality assessment.(DOC)Click here for additional data file.Table S3Multiple comparisons of genotype effects.(DOC)Click here for additional data file.Table S4Stratified analysis of the TLR9 polymorphisms with cancer risk.(DOC)Click here for additional data file.


Deep vein thrombosis is one of the common complications of orthopedic surgery. Studies indicated that genetic factors played a considerable role in the pathogenesis of deep vein thrombosis. Endothelial nitric oxide synthase which encoded by nitric oxide synthase 3 (NOS3), can generate nitric oxide in endothelial cells. As a predominant regulator for vascular homeostasis, nitric oxide might be involved in the pathogenesis of thrombosis. It had been proved that the NOS3 polymorphism (rs1799983) was associated with the development of cardiovascular diseases. Our objective was to evaluate the association between the NOS3 polymorphism (rs1799983) and deep vein thrombosis after orthopedic surgery in Chinese Han population. The polymorphism was genotyped in 224 subjects with deep vein thrombosis after orthopedic surgery and 580 controls. Allele and genotype frequencies were compared between subjects with deep vein thrombosis and control subjects. The allele and genotype frequencies of the NOS3 polymorphism (rs1799983) were significantly different between subjects with deep vein thrombosis and control subjects. There were also significant differences when the subjects were stratified by gender, surgery type and hypertension status. These findings suggested that the NOS3 polymorphism (rs1799983) was associated with susceptibility to the deep vein thrombosis after orthopedic surgery in Chinese Han population, and NOS3 might play a role in the development of deep vein thrombosis after orthopedic surgery. Deep vein thrombosis (DVT) is one of the common complications of orthopedic surgery. It usually occurs at the deep veins of legs  or the deep veins of pelvises after total knee arthroplasty (TKA) and total hip arthroplasty (THA). The incidence of DVT is up to 50–60% in patients after arthroplasty Nitric oxide synthase 3 (NOS3) locates at chromosome 7q36, and it encodes endothelial nitric oxide synthase (eNOS), which can generate nitric oxide (NO) in endothelial cells. Endothelial NO is a key determinant of vascular homeostasis. It can regulate several physiological processes, including platelet adhesion, leukocyte adhesion, endothelial cell migration and vascular smooth muscle cell migration. And it can also participate in vascular repair. Dysfunction of any of these processes may result in atherosclerotic and thrombotic diseases Together, these studies implicated the candidacy of NOS3 as a susceptibility gene for DVT after orthopedic surgery. However, there was no report of association between NOS3 and DVT after orthopedic surgery. The objective of this research was to appraise the genetic association between the polymorphism (rs1799983) in NOS3 gene and DVT after orthopedic surgery in Chinese Han population.Our research was conducted in accordance with the Declaration of Helsinki. This study was approved by the ethics review committee of the Drum Tower Hospital affiliated to Medical School of Nanjing University, and all participants provided their written informed consent to participate in this study.804 consecutive eligible subjects underwent orthopedic surgery on knee or hip were recruited at the Center of Diagnosis and Treatment for Joint Disease, Drum Tower Hospital affiliated to Medical School of Nanjing University. Of these 804 subjects, 224 cases suffered from DVT after orthopedic surgery  and 580 controls showed no evidence of DVT . For each subject, DVT was assessed using venography, only subjects with filling defect of venous lumen were classified as cases, subjects without filling defect of venous lumen were classified as controls, and the healthy controls had no history of VTE or clinical evidence of VTE. Hypertension was defined as systolic blood pressure >140 mmHg and/or diastolic blood pressure >90 mmHg on an average of two measurements or by current anti-hypertensive treatment by WHO  criteria. All subjects enrolled in this study were Han Chinese origin living in or around Nanjing. No subject withdrew from this study.DVT was evaluated by performing venography within 7 days postoperatively for all these 804 subjects. The following venous segments were observed: common femoral veins, superficial femoral veins, anterior and posterior tibial veins, peroneal veins, popliteal veins. The criterion for diagnosis of DVT was a filling defect of venous lumen in two or more views according to the American College of Chest Physicians Evidence-Based Clinical Practice Guidelines of diagnosis of DVT The NucleoSpin Blood QuickPure Kit  was used to extract DNA from peripheral blood following the manufacturer’s protocol. The SNP rs1799983 was genotyped by using Taqman assay , according to the manufacturer’s instruction.p value of <0.05 was considered statistically significant in this study. These tests were implemented by using SPSS 12.0 system software .The previous studies had shown that the G allele played a protective role in coronary artery disease and VTE p>0.05). The Hardy–Weinberg equilibrium of all subjects was tested which indicated there was not genotyping error in our dataset (p  = 0.806). The allele and genotype frequencies of DVT subjects and control subjects were shown in p  = 0.70 and 0.61, respectively) (p  = 0.012) and in the comparison of GG genotype versus other genotypes combined (p  = 0.010) between cases and controls (p  = 0.5265) (The basic characteristics of the study subjects were summarized in ctively) . Significontrols . The sig 0.5265) .p  = 0.001) (p  = 0.000) (p  = 0.926) (p>0.05)   and in t= 0.000) , but no = 0.926) . The sig(p>0.05) .p  = 0.034) (p>0.05) (p  = 0.043) (Significant differences were observed in the comparison of GG genotype versus other genotypes combined (= 0.034)  in hip s(p>0.05)  in knee = 0.043)  in knee p  = 0.001) (p  = 0.001) in the hypertensive group after stratification by hypertension status. The significant differences still existed when adjusted by age, gender and BMI. No significant difference was observed in the normotensive group.Moreover, significant differences were observed in the comparison of allele frequency (= 0.001)  and in tThis study was the first attempt to evaluate the association of the NOS3 polymorphism (rs1799983) with the risk of DVT after orthopedic surgery. On the basis of this study, we demonstrated that there were significant differences in genotype and allele frequencies between DVT subjects and control subjects, the GT+TT genotype and T allele frequencies were significantly higher in cases compared to controls, thus, we stated that the T allele might play a risk role in subjects after orthopedic surgery. To test whether or not the gender, surgery type and hypertension status could impact the results, we stratified the subjects by gender, surgery type and hypertension status, and statistical analyses were performed in these subgroups, we also observed that there were significant differences in genotype and allele frequencies between the cases and controls when Bonferroni’s correction was applied. Our findings were similar to some previous studies which had shown the association of the single nucleotide polymorphism (SNP) in the NOS3 gene (rs1799983) with the risk of coronary artery disease and venous thromboembolism (VTE) Stratification of subjects by gender revealed that significant differences in genotype and allele frequencies between the cases and controls in female subjects, the GT+TT genotype and T allele frequencies were also significantly higher in female cases compared to female controls, it was suggested that the G allele might play a protective role against T allele in female subjects after orthopedic surgery. Whereas, no significant difference was observed in male subjects, and the OR values in the comparison of allele frequency and in the comparison of GG genotype versus other genotypes combined were controversial to those in the comparisons in female subjects. This SNP might be not associated with DVT after orthopedic surgery in male subjects. Association between gender and genotype distribution of rs1799983 were also performed in prior study, the significant differences were observed in male subgroup, and no significant differences were observed in female subgroup Significant differences were observed in the knee surgery group, but no significant differences were observed in the hip surgery group when stratified by surgery type and adjusted by age, gender and BMI. The T allele frequencies were significantly higher in cases compared to controls in the knee surgery group, which suggested that subjects with T allele was prone to DVT after knee surgery. The OR values in the comparison of allele frequency and in the comparison of GG genotype versus other genotypes combined were similar between the two groups, it indicated that the trends of allele frequency and genotype distribution were consistent between the two groups. The explanation of negative results in knee surgery group was the relatively small sample size, which might limit the statistical power. To check the possible association, further researches should be implemented.p value the comparison of TT genotype versus other genotypes combined in the hypertensive group and the p value of all genotypes. The explanation for this discrepancy was the small size of the subjects. Thus, we could not make a definite conclusion because the sample size was small. Further studies with more samples would be necessary to prove the association.Moreover, significant association was observed in the hypertensive group, and no significant association was observed in the normotensive group when stratified by hypertension status. The GT+TT genotype and T allele frequencies were significantly higher in cases compared to controls in the hypertensive group. These results indicated that hypertensive patients with T allele were prone to DVT after orthopedic surgery, and the SNP rs1799983 might be associated with DVT after orthopedic surgery in hypertensive patients. Compared to the normotensive subjects, NOS3 might make more contribution in the etiology of DVT after orthopedic surgery in hypertensive subjects. However, we cannot calculate the In NOS3, rs1799983 locates at c.894 of the coding region, and c.894G/T corresponds to glutamic acid and aspartic acid at position 298 of eNOS protein, respectively. eNOS protein with aspartic acid was unstable compared to that with glutamic acid In conclusion, the present study demonstrated that there might be a significant association between the NOS3 polymorphism (rs1799983) and susceptibility to DVT after orthopedic surgery in Chinese Han population. Further studies would be needed to be conducted to confirm the association of this polymorphism with DVT after orthopedic surgery in Chinese Han population and in other populations.


Previous epidemiologic studies have reported inconsistent results between parity and pancreatic cancer (PC) risk. To our knowledge, a comprehensive and quantitative assessment of this association has not been conducted.Relevant published studies of parity and PC were identified using MEDLINE (PubMed) and Web of Science databases until November 2013. Two authors (H-BG and LW) independently assessed eligibility and extracted data. Eleven prospective and 11 case-control studies reported relative risk (RR) estimates and 95% confidence intervals (CIs) of PC associated with parity. Fixed- and random-effects models were used to estimate the summary RR depending on the heterogeneity of effects.Q = 50.49, P<0.001, I2 = 58.4%). Significant inverse associations were also observed in the studies that adjusted for cigarette smoking , Type 2 diabetes mellitus , and those that included all confounders or important risk factors . Additionally, in the dose-response analysis, the summary RR for per one live birth was 0.97 , which also indicated a borderline statistically significant inverse effect of parity on PC risk. No evidence of publication bias and significant heterogeneity between subgroups were detected by meta-regression analyses.The summary RR for PC comparing the highest versus lowest parity was 0.86 (95% CI: 0.73–1.02; In summary, these findings suggest that higher parity is associated with a decreased risk of PC. Future large consortia or pooled studies are warranted to fully adjust for potential confounders to confirm this association. The search was limited to published studies of humans by using the following search key words and Medical Subject Headings terms: (parity OR pregnancy OR livebirth OR reproductive OR reproduction OR reproductive factors) AND (pancreas OR pancreatic) AND (cancer OR neoplasm OR carcinoma OR tumor). We also reviewed the references of all included studies for additional publications. We followed PRISMA  for conducting and reporting meta-analyses Published studies were included if they 1) used a case-control or prospective study design; 2) evaluated the association between parity and PC risk; and 3) presented hazard ratio (HR), odds ratio (OR), or relative risk (RR) estimates with 95% confidence intervals (CI), standard errors (SE) or data necessary to calculate these. When multiple publications from the same study were available, we used the publication with the largest number of cases and most applicable information. After excluding 1045 and 47 articles based on screening of titles or abstracts, respectively, we identified 27 potentially relevant articles for further full text review  [6]–[32]For each eligible study, two investigators (H-BG and LW) independently performed the eligibility evaluation and data abstraction. The disagreements were discussed and resolved by consensus or by involving a third reviewer (Q-JW) for adjudication. Data abstracted from each study were: author list, year of publication, study region and design, study sample size (number of cases and controls or cohort size), range of follow-up for cohort studies, exposure and outcome assessment including parity categories, study-specific adjusted estimates with their 95% CIs for the highest versus lowest parity, and factors matched by or adjusted for in the design or data analysis. If multiple estimates of the association were available, we abstracted the estimate that adjusted for the most covariates.The study-specific adjusted RRs were used as the measure of association across studies. Because the absolute risk of PC is low, we assumed that estimates of ORs from case-control studies and risk, rate or hazard ratios from prospective studies were all valid estimates of the RR and we therefore report all results as the RR for simplicity. For studies that did not use the category with lowest parity as the reference For the dose-response analysis which calculated parity as a continuous variable, we used the method proposed by Greenland et al. Q statistic and the I2 statistic Q statistic, a P-value less than 0.1 was considered to be representative of statistically significant heterogeneity. I2 represents the proportion of total variation contributed by between-study variation P-value less than 0.05 for meta-regression was considered representative of significant statistical difference between subgroups. Finally, we carried out sensitivity analyses excluding one study at a time to explore whether the results were strongly influenced by a specific study.We evaluated heterogeneity of RRs across studies by using the Cochrane P-value less than 0.05 for Egger's or Begg's tests was considered representative of significant statistical publication bias. Statistical analyses were performed with Stata . P-values were two sided with a significance level of 0.05.Publication bias was evaluated via Egger's linear regression Characteristics of the 22 included articles Of the 11 case-control studies, three were conducted in the United States Q = 50.49, P<0.001, I2  = 58.4%) (P for bias  = 0.652) or with Begg's test (P for bias  = 0.324) and no asymmetry was seen in the funnel plots when inspected visually. In a sensitivity analysis, we sequentially removed one study at a time and re-analyzed the data. The 22 study-specific RRs of parity ranged from a low of 0.84  after omission of the study by Ji et al Q = 45.98, P = 0.001, I2  = 56.5%) after omission of the study by Lucenteforte et al. Eleven prospective = 58.4%) . There wQ = 62.83, P<0.001, I2  = 69.8%)  when the study by Ji et al Q = 56.29, P<0.001, I2  = 68.0%) when the study by Chang et al Q = 43.60, P = 0.001, I2  = 56.4%) which was similar to the original analysis including all studies. There was no evidence of a nonlinear association between parity and PC risk, P for nonlinearity  = 0.2409. Furthermore, when we removed three studies Q = 49.60, P<0.001, I2  = 67.7%) were similar.Nine prospective = 69.8%) . In a seIn subgroup analyses of highest versus lowest categories of parity and PC risk, almost all strata showed inverse associations, although not all of them showed statistical significance. There was no evidence of significant heterogeneity between subgroups with meta-regression analyses . In the To our knowledge, this is the first meta-analysis which provide comprehensive and quantitative evidence of the association between parity and PC risk. In this study, high parity was associated with a borderline statistically significant decreased risk of PC although the decrease in risk was small for each additional live birth. In addition, consistent inverse associations were observed within the analyses stratified by whether the studies included adjustment for cigarette smoking, Type 2 DM, or all of the confounders or important risk factors .Although the exact biological mechanisms by which increased parity may decrease the risk of PC are not well established, a hypothesis that endogenous estrogens may be protective against PC has been proposed based on in-vitro and in-vivo studies Although the results of meta-regression found no evidence of significant heterogeneity between subgroups by study design, the strength of the association estimate for prospective studies was weaker than that from case-control studies although both of them showed inverse associations with PC risk . Given tQ = 65.1, P<0.001) which was observed among the studies did not adjust for type 2 DM, the summary results in these studies adjusted for aforementioned confounders had low heterogeneity. Similar pattern was also observed in the studies whether adjusted for the three confounders   . HoweverThis study had several strengths. To the best of our knowledge, this is the first comprehensive and quantitative assessment of parity and PC. Particular strengths of the current meta-analysis are that it including a total of 8,247 cases and 3,498,673 non-cases which should have provided sufficient statistical power to detect this putative association. A further strength is that we carried out a number of subgroup and sensitivity analyses to explore the potential sources of heterogeneity.Potential limitations of this meta-analysis must be taken into consideration. First, as a meta-analysis of epidemiological studies, this study contains the limitations inherent to combining results from studies with heterogeneous study designs. Cohort studies are less susceptible to bias  than case-control studies because, due to the prospective design, information on exposures is collected before the diagnosis of the disease. Additionally, given the possible differences in detailed confounder adjustment information over time, we also carried out the stratified analysis by published year, but the results were unchanged (data not shown). Second, since we did not have access to the primary data of the studies included in this meta-analysis, we could not perform additional adjustments for potentially important covariates or accurately assign an exposure value to open-ended parity categories In conclusion, this comprehensive meta-analysis provides evidence that increased parity is associated with a slightly decreased risk of PC. Although this relative risk is moderate in size, given the limited established risk factors and low survival rate, further large consortia or pooled studies, are warranted to fully adjust for the potential confounders and focus on the sex-steroid related factors which might play a role in the development of PC malignancies.Table S1Characteristics of studies of parity and pancreatic cancer risk.(DOC)Click here for additional data file.Checklist S1PRISMA 2009 Checklist.(DOC)Click here for additional data file.


The fat mass and obesity associated gene (FTO) polymorphisms have been implicated in the susceptibility of overweight/obesity in children and adolescents. However, the results have been inconsistent. In this study, we performed a meta-analysis to clarify the association of FTO gene polymorphisms with overweight/obesity risk among children and adolescents. PubMed and Embase were used to search for eligible published literatures. Pooled odds ratios (ORs) with 95% confidence intervals (CIs) were calculated using random- or fixed-effect models. P<0.001). The overall pooled ORs for risk obesity and overweight were 1.34 (95%CI: 1.21-1.48) and 1.35 (95%CI: 1.25-1.47), respectively. Subgroup analyses also showed similar trends in most subgroups of adjustment for covariates and unadjustment, different ethnicities , and each of three investigated polymorphisms . A total of 21 articles containing 23 studies (11208cases and 35015controls) were included in our analysis. The results indicated that variant in FTO gene was significantly associated with increased risk of overweight/obesity in children and adolescents (OR=1.35; 95%CI: 1.27-1.44; The present meta-analysis suggested a positive association between FTO gene polymorphism and overweight/obesity risk among children and adolescents. Further prospective studies should be recommended to confirm the observed association, and underlying mechanism should be investigated to clarify the association of FTO gene polymorphism with overweight/obesity. Obesity, which is associated with an increased risk of many chronic diseases, including type-2 diabetes, cardiovascular diseases and cancer, has become a major health problem worldwide . More thFTO, TMEM18, GNPDA2, INSIG2, MC4R, NEGR1, 1q25, BDNF and KCTD15[FTO, a gene located in chromosome region 16q12.2, was the first identified gene for common obesity[FTO variants with BMI and obesity has been investigated in different ethnic populations. A previous meta-analysis by Peng et al. [FTO with overweight/obesity risk in children and adolescents. Obesity is a complex multifactor disease that is affected by environmental risk factors and biological ones (including monogenic variations and common genetic variants). Approxind KCTD15. FTO, a g et al.  indicateFTO) and (polymorphism OR variant OR variation OR genotype) and (children OR adolescents) and (obesity OR overweight). The publication language was restricted to English. The reference lists of retrieved articles were hand-searched. If more than one article were published using the same case series, only the study with largest sample size was included. The literature search was updated on 20 June 2013.We searched the literature databases including PubMed and Embase from 2007 to 2013 since rs9939069 polymorphism in FTO gene and its association with obesity was first reported in 2007. The search strategy to identify all possible studies involved the use of the following keywords:  using case-control, cross-sectional, or cohort design; (3) providing an odds ratio (OR) or relative risk (RR) with 95% confidence interval (CI) under an additive model or sufficient raw data to calculate it; and (4) participants should be children and/or adolescents. The following information was extracted from each study: (1) name of the first author; (2) year of publication; (3) origin of country; (4) ethnicity of studied population; (5) number of cases and controls; (6) mean age and body mass index (BMI) of subjects; (7) OR or RR with 95% CI under an additive model; (8) adjustment for covariates; (9) studied SNPs; (10) genotype distribution in cases and controls; and (11) P value for Hardy-Weinberg equilibrium (HWE) test in controls. Two authors independently assessed the articles for compliance with the inclusion/exclusion criteria, and resolved discrepancies by group discussion until reaching a consistent decision.The included studies fulfilled the following inclusion criteria: (1) an original report evaluating the association of any of the FTO polymorphism and overweight/obesity risk was estimated by calculating pooled OR with 95%CI under an additive model. The significance of pooled OR was determined by Z test . The between-study heterogeneity was evaluated by Q statistic and I2 index[P<0.10) or absence (P>0.10) of heterogeneity, respectively. Subgroup analyses were conducted by ethnicity, adjustment, FTO SNP, and category of cases. Sensitivity analysis, after removing one study at a time, was performed to evaluate the stability of the results. Begg’s funnel plot[P<0.05 was considered statistically significant). Data analysis was performed using STATA version 11.The association between I2 index. A randoI2 index) or fixeI2 index) effectsnnel plot, a scattnnel plotand Eggernnel plot, rs9939609 was considered as the surrogate polymorphism. The ethnicities varied across studies: 16 were on Caucasian population [The genotypes in the controls of all included studies were in Hardy-Weinberg equilibrium, except for two those were unknown ,26 and tPs r2>0.5, rs9939pulation -40,43,44pulation ,30,32,42FTO rs9939609 (or its proxy) with risk of overweight/obesity in children and adolescents, there was evidence of heterogeneity between studies . Therefore, the random effect model was used. The result showed a statistically significant association between rs9939609 and overweight/obesity, with an overall OR of 1.35 . The overall pooled OR of risk overweight was 1.35. In the stratified analysis by adjustment, the pooled summary OR with the adjusted data showed significant association , and the overall analysis without adjusted data also showed the similar result . Further subgroup analysis stratified by ethnicity showed significant association between rs9939609 and overweight/obesity risk in Caucasians , in Asians , and in Amerindians , but not in Africans  . In addition, we also performed a stratified analysis according to different polymorphisms. Except for rs8050136 , all the polymorphisms showed a strong correlation with increased risk of being overweight/obese . The results of subgroup analyses were showed in Regarding to the association of P for HWE was more than 0.01, the results did not substantially change, with ORs ranging from 1.30(95%CI:1.25-1.35) to 1.38(95%CI:1.33-1.44). Thus, the significant association for rs9939609 with overweight/obesity risk was statistically robust by sensitivity analysis.To test the sensitivity of the meta-analysis, we excluded each study at one time. The corresponding pooled ORs were not materially altered. After excluding the two studies where the HWE of the genotypes in controls were unknown, and two studies where P=0.064; Egger’s test: P=0.637). Therefore, no publication bias was detected in this mate-analysis. The Begg’s funnel plot did not reveal any evidence of obvious asymmetry , and botIn this study, our meta-analysis confirmed that variant in FTO gene was significantly associated with increased risk of overweight/obesity in children and adolescents. The stability of sensitivity analysis and no publication bias confirmed the positive association. Further subgroup analyses indicated the similar trends among subgroups of overweight and obesity, data with and without adjustment, different ethnicities , and each of three polymorphisms investigated .Between-study heterogeneity is usually common in meta-analysis of genetic association study. As the FTO contributes to the increased risk of obesity is largely unknown[FTO plays an important role in energy homeostasis, metabolism and adipogenesis [FTO variant affects fat mass may help us to understand the pathogenesis of obesity[FTO was influenced by higher fatty acid intake[FTO variants on pituitary function has a strong association with TSH levels, for both of them are expressed or produced in pituitary gland[FTO genetic effects do not begin to influence early onset obesity before the age of 7 years[FTO in childhood may be effective in identifying individuals genetically predisposed to obesity, the lifestyle factors also should be considered [The underlying biological mechanism on how the variant in y unknown. For aniogenesis ,47. To ef obesity. FTO genf obesity and dimif obesity . Meanwhid intake,. Some rid intake,49. Besiid intake. The FTOid intake. Regardiid intake, influenary gland. Howeverf 7 years. Despitensidered . FurtherMC4R gene, another obesity susceptibility gene, in children reported by Xi et al. [The result of pooled effect for rs9939609 genetic variant of FTO gene with overweight/obesity risk in our meta-analysis  was similar with previous overall result of rs9939609 by Peng et al. [.19-1.33). However.19-1.33). Moreove.19-1.33). FurtherDespite the many strengths compared to individual studies, there are several limitation in our meta-analysis. First, some potential confounding factors were not controlled for because our pooled estimate was based primarily on unadjusted estimates and CIs. Second, different criteria of obesity and overweight were included in our meta-analysis. Therefore, these results should be interpreted with caution. Trying to overcome this drawback, we performed subgroup analysis by ethnicity, which indirectly reflected the differences in obesity and overweight criteria as the cut-offs were similar in the same ethnic group. Third, a number of obesity-related clinical and biochemical parameters were not estimated in this meta-analysis. Forth, owing to insufficient original data from the included studies, the effects of gene-gene and gene-environment interactions were not assessed. In summary, our meta-analysis results indicated that rs9939609 polymorphism in the FTO gene was significantly associated with overweight/obesity risk in children and adolescents. However, further studies considering gene-gene and gene-environment interaction should be conducted to investigate the association. Furthermore, better estimates of the risk would be obtained in future prospective cohort studies, as well as more mechanism researches are also needed to clarify the association of FTO gene with overweight/obesity. Checklist S1PRISMA checklist.(DOC)Click here for additional data file.


MEF2A) gene, the first identified CAD-causing gene, has attracted special attention but the results are controversial. We aimed to evaluate this genetic association via a case-control study and meta-analysis.Coronary artery disease (CAD) is the most common heart disease worldwide. Association of CAD with variants in the myocyte enhancer factor 2A . No significant link was observed between the common (CAG)n polymorphism and CAD, whereas the rare 21-bp deletion was detected only in five affected individuals. The meta-analysis of (CAG)n polymorphism and CAD risk, including nine studies with 3801 CAD patients and 4020 controls, also provided no convincing evidence for the genetic association, even upon stratification by race (mainly Whites and Chinese). However, the 21-bp deletion was regarded as a potentially logical, albeit undetermined, candidate for CAD in the following systematic review.We performed a case-control association study to investigate the relationship between variations in exon 11 of n polymorphism with CAD, however, we concluded that the rare 21-bp deletion might have a more compelling effect on CAD than the common (CAG)n polymorphism, and MEF2A genetic variant might be a rare but specific cause of CAD/MI.Our findings failed to demonstrate a correlation between (CAG) Coronary artery disease (CAD) is a common complex disorder resulting from both genetic and environmental influences MEF2A) gene, was reported as a causative mutation in a single large CAD/MI family of Scandinavia ancestry In vitro functional analysis indicated that the 21-bp deletion disrupted the nuclear localization of mature protein and decreased MEF2A-induced transcriptional activation. Thus this genetic imperfection might lead to a defective or abnormal vascular endothelium, which could promote the genesis of atherosclerotic plaque or thrombosis and influence the whole process of atherogenesis MEF2A gene has been considered as the first CAD-causing gene to be identified.In 2003, a 7-amino acid deletion, caused by a 21-base pair (bp) coding sequence deletion in exon 11 of the myocyte enhancer factor 2A  polymorphisms such as a common variant (CAG)n polymorphism. However, the results have been inconsistent The genomic sequence of MEF2A locus on chromosome 15q26 nor its adjacent region has been identified in any of the previous GWASs, thus generating debate over the nature of MEF2A genetic contribution to individual susceptibility to CAD.With the improved genotyping technologies and the completion of the human HapMap project, Genome-Wide Association Studies (GWASs) have been developed as an important approach in genetic research. Thus far, a large number of candidate loci conferring risk of or protection from common complex diseases such as CAD have been proposed MEF2A gene and its effect on CAD risk, we focused on its exon 11, the highly polymorphic and controversial region, and established a well-characterized case-control study of 1045 sporadic CAD patients and 1008 controls with normal coronary arteries. In addition, we reviewed all available studies reported in the literature to examine the association of the common (CAG)n polymorphism and the rare 21-bp deletion with CAD, and to assess whether variations in study design and study population ethnicity could lead to potential biases and be the sources of between-study heterogeneity.To elucidate the relationship between Approval to undertake this study was granted by the Ethics Review Committee of Ruijin Hospital, Shanghai Jiao Tong University School of Medicine and was conducted according to the Declaration of Helsinki Principles. Written informed consents were obtained from each participant at enrollment.This was a hospital-based case-control study including a total of 2053 unrelated Han Chinese admitted to Ruijin Hospital, Shanghai Jiao Tong University School of Medicine when they were experiencing various symptoms or for a medical checkup from January 2006 to September 2009. All participants underwent coronary angiography and were divided into CAD group and control group according to their angiographic results.The CAD group contained 1045 sporadic patients aged 65.49±9.83 years, and the diagnosis of CAD was determined angiographically based on the presence of more than 70% stenosis in at least one of the three major coronary arteries or major branches. Patients with simple spasm of coronary arteries, myocardial bridge or other non-coronary atherosclerotic lesions were excluded. The remaining participants (n = 1008), aged 60.23±10.49 years, had normal coronary arteries (NCA) on angiography, formed the control group.All patients with the 21-bp deletion were followed up every year in a special CAD clinic. At each visit, clinical manifestations and echocardiography were recorded. Adverse events  were reported during the visit or through telephone conversation with the patients or their family members. Two trained physicians independently reviewed all medical notes, including emergency department visit forms and hospital medical records.MEF2A exon 11 in this cohort study, we sequenced the entire exon 11 using the direct DNA sequencing method in all 2053 subjects. Primers were designed by the Primer3 software (http://frodo.wi.mit.edu/cgi-bin/primer3/primer3) according to reference sequence (NM_005920.2). In detail, the sequence of the forward primer was 5′-gca gag gta ctt gca agc cat ctg-3′ and the reverse was 5′-ggt cgg cca agc aca att gga gaa-3′. The sequencing primer was 5′-caa gca caa ttg gag aat gga-3′. Sequences were analyzed using an ABI Prism BigDye Terminator Cycle Sequencing Kit on an ABI Prism 3700 sequencer, version 3.1 , as described in detail in Blood samples (5 ml) were drawn and genomic DNA was extracted from peripheral blood leukocytes by standard phenol-chloroform extraction. To assess the distribution patterns of the structural variations of viz. PubMed and Excerpta Medica Database (EMBASE), and two Chinese searching engines, viz. Wanfang database (http://www.wanfangdata.com.cn) and China Biological Medicine (CBM) (http://sinomed.imicams.ac.cn/index.jsp) with the last update on July 31, 2011. We restricted search results to papers published in English or Chinese. We combined the subject terms of ‘coronary artery disease or coronary disease or arteriosclerosis or atherosclerosis or myocardial infarction or angina pectoris’ and ‘myocyte enhancer factor 2A’ with either ‘gene’, ‘variation’, ‘variant’, ‘mutation’, ‘polymorphism’ or ‘allele’, which were all MeSH  terms. The “related articles” in the MEDLINE option as well as reference lists of all retrieved studies were also checked for citations of other relevant publications that were not identified initially. All studies were considered potentially eligible if they aimed to investigate the relationship between MEF2A genetic polymorphisms and CAD risk. If there were multiple publications from the same study group, the most complete and recent results were extracted. Search results were limited to studies performed in human subjects without country restrictions and ethnic restrictions.We collected information via two international searching engines, MEF2A genetic polymorphisms with CAD/MI, 2) genotyping had been performed by using validated methods, and 3) they provided the sufficient information on genotype/allele counts or frequencies for estimating odds ratio (OR) and 95% confidence interval (95% CI). We calculated the effect estimate against healthy subjects/NCA controls.We enrolled all prevalent case-control or nested case-control or cross-sectional studies in this meta-analysis regardless of sample size, if 1) they explored the association of Data were extracted independently and entered into separate databases by two authors (Y. Liu and W. Niu) from each qualified study: first author's last name, publication date, population ethnicity, study design, diagnostic criteria, genotyping methods, baseline characteristics of the study population, such as age, gender, history of hypertension and diabetes mellitus, if available, and the number of persons with different alleles in cases and controls and available subgroups. Discrepancies between the two databases were identified by comparison. A third author (W. Jin) checked for them and a consensus was reached after discussion. For consistency, continuous variables such as age were uniformly expressed as mean ± standard deviation (S.D.)t-test for continuous variables and the 2χ-test for categorical ones were used to test differences between cases and controls, OR of CAD risk and their 95% CI were calculated as well.For our case-control study, database management and statistical calculation were conducted using SPSS version 13.0 . The Student's http://anthro.unige.ch/software/arlequin). The Haplo.stats package (version 1.4.0) in the R statistical computing software (http://www.r-project.org) was used to analyze haplotype-based association study. Two-tailed P<0.05 was accepted as statistically significant.Hardy-Weinberg equilibrium calculations were performed with the Arlequin program . Using the most common type (CAG)9 allele as a reference, comparisons of other (CAG)n alleles between cases and controls were expressed in the form of OR and 95% CI. The allele effects were estimated using the model-free approach, where no assumption about genetic models was required. In addition, stratification analyses were conducted to seek more narrowly drawn subsets of the studies such as different genotyping methods, population origins and study designs. We implemented the random-effects model using the method of DerSimonian and Laird, instead of fixed-effects model, to bring the individual effect-size estimates together, and the estimate of heterogeneity was analyzed by the Mantel-Haenszel method In the following meta-analysis, pooled association relating (CAG)2χ-based Cochrane's Q statistic with statistical significance at the level of 0.10 as this statistic has proven to have poor power if there are few studies 2I statistic was documented for the percentage of the observed between-study variability due to heterogeneity rather than chance with the ranges of 0–100%  The presence of between-study heterogeneity across all eligible comparisons was calculated using the fsN) of each meta-analysis fsN was smaller than the number of observed studies for a polymorphism, it is believed that the meta-result might have a significant publication bias. In this study, the fsN significance was established at P<0.05 (fs0.05N = (ΣZ/1.64)2−k; where k is the number of articles included in each meta-analysis).Finally, publication bias was assessed by the fail-safe number (P<0.001) and more often of the male gender (P<0.001). As expected, the CAD group had a higher prevalence of conventional cardiovascular risk factors, including diabetes and dyslipidemia (P<0.05). They had higher serum levels of fasting glucose, total cholesterol, triglycerides, and low density lipoprotein-cholesterol, and lower levels of high density lipoprotein-cholesterol. However, the morbidity of hypertension was similar between the two groups.The clinical characteristics of our study population are shown in Eight variants were identified by sequencing the entire exon 11 in 2053 unrelated Chinese individuals . No signn(Q)) spanned from 4 to15, and the majority of individuals had 9–11 repeats. Shown in n polymorphism. No statistical significance was observed (P = 0.347) for the allelic association of this polymorphism with CAD, and the distribution of genotypes was also similar in two groups (data not shown).The number of the CAG triplet repeats . More than 95% of individuals in both CAD patients and NCA controls contained five prolines, and the frequencies of (CCG)n allele and genotypes yielded no significant differences between two groups (data not shown). Additionally, a CCA deletion resulting in lack of one proline amino acid, located adjacent to (CCG)n site, was detected only in one unaffected subject.Closely following the (CAG)Interestingly, the 21-bp deletion was found only in five independent CAD patients, and none in NCA subjects. In this cohort, they all had some traditional CAD risk factors, including dyslipidemia, hypertension and family history of cardiovascular diseases; three showed severe lesion in the left main coronary artery and two were diagnosed with premature CAD. After a 5-year follow up, one died of sudden cardiac death, one took stent treatment and three underwent coronary artery bypass grafting .Besides the aforementioned four variants, we identified three synonymous SNPs  and one non-synonymous SNP (C1303T) in exon 11. The A1299G and C1303T polymorphisms were rare SNPs with minor allele frequencies being 0.1% and 0.4% among NCA controls, respectively. As a result, none of these four SNPs were associated with CAD (data not shown).9-5P-G-T (in order of (CAG)n, (CCG)n, G1305A and G1353T) (26.0% in NCA controls) as the baseline for the comparison of the rest, the differences in haplotype distributions between the two groups did not achieve nominal significance (data not shown).After dropping four rare variations , we evaluated the remaining four common variants for further haplotype analysis. Using the most common haplotype (Q)MEF2A gene variations on CAD was evaluated. A total of fifteen studies met selection criteria, whereas only eight studies n genotypes/alleles, and one shared the same population The initial search strategy retrieved forty-three relevant articles in English (n = 24) and Chinese (n = 19), in which the effect of n polymorphism except for two with only genotype counts 9 allele frequency differed widely in diverse ethnic groups. In Whites, the frequencies were in the ranges of 32.2% to 36.2% for CAD cases and 30.6% to 36.3% for controls, which were lower than that in Chinese ranging from 24.3% to 40.5% for cases and 22.7% to 41.8% for controls. In contrast, the Turks had a higher frequency of 51% for cases and 33.5% for controls  under a random-effects model. Whereas, of nine studies, only two individual OR estimates showed a higher risk of CAD that was statistically significant for (CAG)9 allele compared with other alleles .As shown in heterogeneityP = 0.25). In comparison, studies in PCR-SSCP group were heterogeneous . However, differences in MEF2A genotyping methods did not affect the overall results materially.However, statistically significant heterogeneity was evident in most subgroups, according to covariates identified by our qualitative assessment . In viewheterogeneityP = 0.01 and <0.00001, respectively). Moreover, negative associations persisted across all comparisons.After stratification by control selection criteria based on clinical symptoms or coronary angiographic data, significant heterogeneity was observed in both the NCA group and the symptom investigation group (heterogeneityP = 0.62), it was significant in Chinese population possibly due to the wide spectrum of (CAG)9 allele . Since there was only one study performed in Turks with a relatively small sample size (n = 156), the risk estimate showed a significant higher risk of (CAG)9 allele with CAD (P = 0.002), and there was no difference in the pooled risk estimates.We divided the population into three groups by ethnicity, Chinese (including this study) fs) at the level of 0.05 for each comparison. The Nfs0.05 values for all the comparisons were greater (62.63) than the number of studies (n = 9) included in this meta-analysis. Therefore, no evidence showed publication bias for association between MEF2A gene (CAG)n polymorphism and CAD susceptibility.To assess publication bias, we calculated the fail safe number n polymorphism, while the other seven were all downstream of this polymorphism within 100 bp. Such intense variation in the context of a single exon led us to explore the link of MEF2A genetic polymorphisms to CAD/MI. A possible explanation might be the remarkable diversity embedded in (CAG)n polymorphism. We therefore carried out a rigorously-designed case-control association study focusing on MEF2A exon 11 in southern Chinese and reviewed all available information regarding the relationship between this genetic hotspot and sporadic CAD/MI from the literature. To the best of our knowledge, this is the first meta-analysis seeking to clarify the association of MEF2A gene (CAG)n polymorphism with CAD risk.In the present study, we verified eight variants in n polymorphism and CAD susceptibility, even upon stratification by race (mainly Whites and Chinese). This meta-analysis had sufficient statistical power n polymorphism itself exhibits null association with CAD, or its effect on CAD is small and depends on neighboring variants that compensate or dilute the variation under study.Data from our case-control study, which was in concordance with most previous observations, and in combination with all other eight studies involving a total of 3801 CAD patients and 4020 controls, failed to show a significant association between (CAG)9 allele with an increased risk and severity of CAD, while data from Dai et al. n polymorphism; the phenomenon was not compatible with the situation expected from random mating. After separating analyses by genotyping methods, we found that heterogeneity between studies in the PCR-SSCP group was higher than the overall estimate; however, there was no indication attributable to the diversity between different experimental methods. Nevertheless, applying appropriate genotype techniques remain an open question.Noteworthy, one study conducted by Han et al. n polymorphism might not be a major contributing locus or have limited values to assess an exact role of MEF2A in CAD/MI, we maintain that application of coronary angiographic criteria for controls is preferable, and the proper phenotype discrimination is critical in any genetic association study.Moreover, it is well known that CAD is frequently asymptomatic and the diagnosis relies on coronary angiography. However, the definition of controls was debatable in a number of the available studies. Some controls were enrolled according to their clinical symptoms and should have been properly defined as “uncertain phenotype” n repeats across different populations. Specifically, the high versus low frequency of (CAG)9 allele was nearly double in both CAD patients and controls, suggesting a possible role of differences in genetic background and the environment in which the populations live. Of note, there was only one eligible Turkish population 9 allele and CAD in the Turkish study might be due to chance or confounding, for its sample size was rather small (n = 156) and its deviation would have little or no effect on our null results. We agree that more studies in diverse ethnic/racial groups are required to draw a firm conclusion.Meanwhile, we observed the wide divergence of (CAG)n polymorphism, cannot be ruled out. The challenge here is to decide which observed variants in MEF2A gene could be considered as a susceptibility or causal mutation in CAD. Our data indicate that the rare 21-bp deletion might have a more compelling effect on CAD than the common (CAG)n variant, and MEF2A genetic variants might, therefore, be a rare but specific cause of CAD/MI. Needless to say, a composite effect encompassing all influential MEF2A genetic variants remains to be determined.It was noteworthy that we identified the 21-bp deletion only in affected individuals, which was consistent with the results from Wang et al. MEF2A gene and others that operate in the same pathway, the single-locus based case-control study and meta-analysis preclude the possibility of gene-gene and gene-environment interactions and may not reveal the full picture. Although there was no evidence showing publication bias in our overall meta-analysis, considering the above limitations, further studies with larger sample size and different ethnic compositions, which typically considered as small or moderate effects, are warranted to avoid study bias.Finally, some limitations of this study should be acknowledged when interpreting the results. Firstly, it is recognized that differences in study design, genetic heterogeneity and statistical methods made it harder to estimate the exact underlying genetic contribution to disease susceptibility. Moreover, some large-scale studies MEF2A gene (CAG)n polymorphism in CAD. However, we suggested that the 21-bp deletion might be a rare but specific cause of CAD. As few studies are available in this field and current evidence remains limited, this conclusion requires further confirmation by well-designed prospective studies with adequate methodological quality and properly controlling for possible confounds, particularly different genetic approaches, homogeneous CAD patients and well-matched controls, gene-gene and gene-environment interactions, and multiethnic groups.In conclusion, our case-control study and the following meta-analysis provide no convincing evidence for the genetic involvement of Text S1Sequence analysis.(DOC)Click here for additional data file.Table S1The clinical characteristics of our study population.(DOC)Click here for additional data file.Table S2Demographics of the CAD patients with 21-bp deletion in our study population.(DOC)Click here for additional data file.


Fixed and random effect models were built to evaluate the association of the four polymorphisms with the risk of tuberculosis, and the association between the four polymorphisms and tuberculosis was expressed as the odds ratio (OR) and 95% confidence interval (CI). Finally, twenty nine qualified studies were enrolled for this meta-analysis that included 6179 tuberculosis cases and 6585 healthy controls. The variant homozygote genotype of the FokI polymorphism was associated with a significantly increased risk of tuberculosis when compared to the heterozygote and wild type homozygote genotypes in the Chinese population . For European subjects, the homozygote and heterozygote genotypes of the BsmI polymorphism were associated with a significantly decreased risk of tuberculosis when compared to the wild type homozygote . Based on the above results, we conclude that variants of the VDR gene that are homozygous for the FokI polymorphism might be more susceptible to tuberculosis in Chinese. Furthermore, larger sample studies are warranted to confirm the protective effects of BsmI variants on tuberculosis in the Europeans.The relationship of four potentially functional polymorphisms of the vitamin D receptor (VDR) gene, ApaI, BsmI, FokI and TaqI , with tuberculosis susceptibility were considered. The aim of this meta-analysis was to explore the association between the four polymorphisms and tuberculosis risk in different ethnic backgrounds. Eligible case-control studies that were catalogued before April 1 In recent years, the anti-bacteria effects of Vitamin D in the human immune system have been explored extensively . MeanwhiDue to underpowered design or small study sample size, previous studies have not reached a consensus regarding the association between the VDR gene variants and tuberculosis risk -11. A prth 2013 to April 1st 2013, and all the searched results were finally updated on April 1st 2013. All articles catalogued before April 1st 2013 were considered. Two medical subject headings (Mesh), “Tuberculosis” and “Vitamin D receptor”, were used to search for related articles, and qualified articles were included according to the following criteria: (1) the original articles were published in English; (2) the studies had a case-control design and were conducted in human beings; and (3) the articles contained available genotyping information on VDR gene polymorphisms . Two independent investigators were employed to search the related articles according to the above criteria.We searched two databases for association studies of VDR gene polymorphisms and tuberculosis: PubMed and Web of Science. We conducted the search from March 18The two investigators independently extracted data and reached a consensus on all of the items. If the two investigators generated different results, they would check the data again and finally reach a consensus. An expert would be invited to the discussion if they could not reach an agreement. Data extracted from the selected articles included the first author’s name, year of publication, country of origin, ethnicity, number of cases and controls, diagnosis method of cases and the selection of controls. 2-test-based Q statistic test was performed to assess the between study heterogeneity, and P<0.05 was considered a significant level. A fixed-effect model by the Mantel–Haenszel method and a random-effect model by the DerSimonian and Laird method were adopted for combining data. When the P value of heterogeneity was <0.05, then a random-effect model was employed. Otherwise, the fixed-effect model was adopted throughout the analyses. These two models provided similar results when heterogeneity between studies was absent; otherwise, a random-effect model was more appropriate. Bonferroni correction was adopted for multiple comparisons.The association between the four polymorphisms and risk of tuberculosis for each study was estimated by the odds ratio (OR) and 95% confidence interval (CI). A χHeterozygote and variant homozygote of each SNP was compared with the wild-type homozygote to estimate the risk of tuberculosis. Then, a dominant model, that combined variant homozygote and heterozygote genotypes, was established to calculate the risk of tuberculosis, versus the wild-type homozygote. Meanwhile, heterozygote and wild-type homozygote genotypes of each SNP were also combined as a recessive model to evaluate the risk of tuberculosis, versus the variant homozygote.Stratification analyses were performed based on ethnicity and region, and a meta regression was used to illustrate the potential reasons for heterogeneity between the studies. Inverted funnel plots, an Egger’s test and a Horbold-Egger’s test were used to assess publication bias . Meta-anA flow chart of the inclusion and exclusion procedures of the articles is shown in In Of the seventeen studies that were conducted in Asia, two were form China, one was from Cambodia, and the remainder originated from India and Iran. Because the geographical differences might contribute to the genetic diversity, we classified the studies into two groups: East and Southeast Asia (China and Cambodia) and South and West Asia (India and Iran). Additionally, seven African studies were conducted in the sub-Saharan region. As a result, we classified the enrolled studies into five subgroups based on ethnicity and region for the subgroup analysis. The five subgroups included East and Southeast Asians, South and West Asians, Africans, Europeans and Americans.recessive=1.36, 95%CI: 1.14-1.62, Pbonferroni=0.0028; heterogeneity test: χ2=33.28, P=0.06)  , and the dominant model of the FokI polymorphism was also significantly increased tuberculosis risk by 0.81 fold . The protective effect of heterozygote of the FokI polymorphism was in marginal significance in South and West Asians (P=0.04). The forest plots of the VDR gene FokI polymorphism (recessive model) in South and West Asians, Africans and Americans showed no significant associations  . Further P=0.62) . Also inciations , 5 and 6 . FurtheThe other three SNPs of the VDR gene showed no notable associations with tuberculosis risk in the pooled samples. However, stratification of ethnicities revealed some evidence of tuberculosis risk for the other three SNPs. dominant=0.41, 95%CI: 0.22-0.76, Pbonferroni=0.02; heterogeneity test: χ2=2.59, P=0.11). Although the heterozygote of the BsmI polymorphism decreased the risk of tuberculosis by 0.53-fold when compared to the wild type homozygote , the Bonferroni correction showed no significance. For the BsmI polymorphism in the European population, the variant homozygote and heterozygote genotypes were associated with a significantly decreased risk of tuberculosis when compared to the wild type homozygote . Also, the heterozygote of the TaqI polymorphism reduced the risk of tuberculosis by 0.11 fold when compared to the wild type homozygote . However, the adjusted P-values of the above results showed no significance by Bonferroni correction.For the TaqI polymorphism in the African population, the variant homozygote and heterozygote genotypes reduced the risk of tuberculosis when compared to the wild type homozygote by marginal significance  exceeded 0.05. Therefore, we did not perform further meta-regression.Meta regression was adopted to illustrate the potential reasons for heterogeneity between the studies if the t=0.51, P=0.614) showed no evidence of publication bias. However, the funnel plot of the FokI recessive model suggested some asymmetry that might be induced by small sample sizes .In this meta-analysis, a Funnel plot, Egger’s test and Horbold-Egger’s test was used to evaluate the potential publication bias of the enrolled studies. For the significant result of the FokI recessive model, the Egger’s test  significantly increased the risk of tuberculosis, and the risk effect was found in the Chinese population. The other three SNPs variants were not linked to any significant risk or protective effect on tuberculosis in the total population. However, in the stratification of different ethnicities, we found that the dominant model of BsmI SNP generated a protective effect on tuberculosis in the European population. http://hapmap.ncbi.nlm.nih.gov/), the four SNPs are valuable for tuberculosis susceptibility research in different ethnic backgrounds. Genetic alterations of the VDR gene may lead to changes in gene activation or in VDR protein structure, and these changes could affect the cellular function of 1,25-dihydroxyvitamin D3. Additionally, VDR gene polymorphisms could be linked to each other and to other unidentified genes, which could also be important for tuberculosis risk.Because the frequency of minor alleles of each SNP of VDR gene was commonly more than 0.01 in the different populations based on the HapMap data (http://hapmap.ncbi.nlm.nih.gov/). Also, due to the small sample size, the analysis of the effect of the FokI polymorphism on tuberculosis was underpowered.The FokI SNP was located within the exon 2 of the VDR gene, and evidence of FokI functionality has already been obtained . ResultsThe ApaI and BsmI polymorphisms were located in the intron region between exon 8 and exon 9 of the VDR gene. Although the nucleotide changes of the ApaI and BsmI polymorphisms generated no changes in the amino acid or the structure of the expressed VDR protein, they might be in linkage disequilibrium with other functional polymorphisms which regulated VDR gene expression. Earlier studies have provided evidence of differential luciferase activity for the two 3' UTR variants that were linked to the most frequent haplotypes . In thisThe allele changes of the TaqI SNP would not generate an amino acid transformation. The t allele of the TaqI SNP seemed to be protective for tuberculosis in the African population, although the results were only marginally significant. This different might be explained by different ethnic backgrounds. The t allele frequency (HapMap data) of TaqI was 0.012 in the Chinese population and 0.118 in the Japanese population, 0.288 in the African population and 0.438 in the European population. Meanwhile, It has been suggested that the mRNA coded from the TaqI t allele of the VDR gene would be more stable than the mRNA from the T allele of the VDR gene , which mMycobacterium tuberculosis and non-tuberculosis Mycobacterium , which might exert a confounding effect.Finally, some limitations of this study need to be addressed. First, HIV status might influence tuberculosis incidence; so, the stratification of HIV status would further reveal the relationship between VDR gene SNPs and tuberculosis. However, HIV status was not reported in one-third of the enrolled studies. Therefore, it was not possible to apply stratification according to HIV status. Second, subgroup analysis with a small number of studies would reduce the statistical power of the analysis. Third, the main method for tuberculosis diagnosis was based on bacteriology. However, these bacteriological methods could not distinguish between In conclusion, this meta-analysis study evaluated the relationship between the VDR gene hot spot SNPs and the risk of tuberculosis in twenty-nine case-control studies, and provided evidence that the FokI SNP might increase the risk of tuberculosis in a recessive model. Thus, host genetic susceptibility might be involved in tuberculosis development. However, due to the small sample sizes, conclusions could not be drawn regarding the putative protective effect of the BsmI variant allele in the European population. Studies with larger sample sizes studies are warranted to be undertaken.Checklist S1PRISMA Checklist.(DOC)Click here for additional data file.Figure S1Forest plot of the dominant model of VDR gene ApaI polymorphism.(PDF)Click here for additional data file.Figure S2Forest plot of the recessive model of VDR gene ApaI polymorphism.(PDF)Click here for additional data file.Figure S3Forest plot of the dominant model of VDR gene BsmI polymorphism.(PDF)Click here for additional data file.Figure S4Forest plot of the recessive model of VDR gene BsmIpolymorphism.(PDF)Click here for additional data file.Figure S5Forest plot of the dominant model of VDR gene FokIpolymorphism.(PDF)Click here for additional data file.Figure S6Forest plot of the dominant model of VDR gene TaqI polymorphism.(PDF)Click here for additional data file.Figure S7Forest plot of the recessive model of VDR gene TaqI polymorphism.(PDF)Click here for additional data file.