To comprehensively evaluate the association of ERCC1 C8092A and ERCC2 Lys751Gln polymorphisms with the risk of glioma.Potential studies were searched and selected through the Pubmed/MEDLINE, EMBASE, the China National Knowledge Infrastructure (CNKI) platforms, WanFang and VIP database up to June 2013. Two investigators independently reviewed full text and included studies met inclusion criteria. Combined odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated in a fixed-effects model or a random-effects model according to results of heterogeneity test. All analyses were performed by Revman 5.2 and Stata 10.0 software.P = 0.01; recessive model: OR = 1.29; 95% CI: 1.07–1.55, P = 0.01). When stratified by ethnicity, significant association was only observed in the Chinese population . For ERCC2 Lys751Gln polymorphism, no significant association was found between ERCC2 Lys751Gln polymorphism and the risk of glioma in different genetic models. A significant association of ERCC2 Lys751Gln polymorphism with the risk of glioma was identified in the Caucasian population under recessive model , but not in the Chinese population.A total of 10 studies were included in our meta-analysis, including 3,580 glioma patients and 4,728 controls. Overall, ERCC1 C8092A polymorphism was associated with the risk of glioma (AA vs. CC: OR = 1.29, 95%CI: 1.07–1.55, This meta-analysis suggested that the AA genotype of ERCC1 C8092A polymorphism might increase the susceptibility of glioma in the Chinese population. And the TT genotype of ERCC2 Lys751Gln polymorphism may decrease the risk of glioma in the Caucasian population. But the small number of studies and moderate methodological quality require cautious interpretation of the study results. Glioma is one of the most common brain tumors, accounting for approximately 80% of all brain tumors Like other cancers, the development and progression of glioma are also determined by genetic and environmental factors. Currently, there are several confirmed environmental risk factors, including ionizing radiation, ultraviolet (UV) rays, diet and smoking. It is evidenced that the low exposure of ionizing radiation and ultraviolet (UV) rays would reduce the susceptibility of glioma 2 = 6.015, P = 0.014) Several single nucleotide polymorphisms (SNPs) have been identified in ERCC1 and ERCC2 genes. But ERCC1 C8092A and ERCC2 Lys751Gln polymorphisms have significant influences on gene expression and DNA repair functions. ERCC1 C8092A polymorphism, located in the 3′-untranslated region of the ERCC1 gene, has been shown to influence the level of ERCC1 mRNA expression and may affect mRNA stability for ERCC1 Evidently, numerous studies have been done to identify the association of ERCC1 C8092A and ERCC2 Lys751Gln polymorphisms with the risk of glioma in different ethnic populations. But the findings were not consistent. In this study, we performed a meta-analysis of the available studies in different ethnic populations to evaluate the effects of ERCC1 C8092A and ERCC2 Lys751Gln polymorphisms on the susceptibility to glioma.We searched for relevant studies up to June 2013 in both English and Chinese through the Pubmed/MEDLINE, EMBASE, the China National Knowledge Infrastructure (CNKI) platforms, WanFang and VIP database with the following terms and their combinations: “ERCC1”, “ERCC2 or XPD” or “ERCC”, “glioma” and “polymorphism or variant”. We tried to identify potential relevant studies from the whole reference lists by orderly reviewing title, abstract and full text.The inclusion criteria were as follows: a). Research focused on the association of ERCC1 C8092A (rs3212986) or ERCC2 Lys751Gln (rs13181) polymorphisms with the risk of glioma; b). Case-control studies; c). Genotype and allele data available. Studies were excluded for following reasons: a). unpublished papers, dissertations, conference articles, reviews and duplication of publications (select the study in a larger sample size); b). data unavailable for calculating genotype or allele frequencies; c). genotype distribution of control subjects violates the Hardy–Weinberg equilibrium (HWE).All the following information was separately extracted by two investigators, including: the first author, year of publication, country (ethnicity), allele frequencies and genotype distributions in glioma cases and controls, number of glioma cases and controls. Extracted data were compared and cross referenced. Inconsistencies were discussed and corrected together.We evaluated the methodological quality of included studies using the Strengthening the Reporting of Genetic Association studies (STREGA) recommendations, extended the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) checklist I2 statistic for estimation of inconsistency was analyzed using the heterogeneity Q statistic test. The following cut-off points indicated different degrees of heterogeneity: I2 = 0–25%, no heterogeneity; I2 = 25–50%, moderate heterogeneity; I2 = 50–75%, large heterogeneity; I2 = 75–100%, extreme heterogeneity P ≥ 0.10), the method of Mantel-Haenszel was used to calculate the pooled OR (95%CI) in a fixed effects model. Otherwise, the DerSimonian-Laird method was performed for evaluating the pooled OR (95%CI) in a random effects model. Begg's funnel plot and Egger's test were performed to assess publication bias among the literatures by Stata 10 software. Funnel plots were used to evaluate publication bias. P<0.05 was considered statistically significant.All meta-analysis was performed by Review manager 5.2 software (The Cochrane Information Management System). The pooled odds ratios (OR) and 95% confidence interval (CI) were calculated for measuring the genetic association between ERCC1 or ERCC2 polymorphisms and the risk of glioma. The between-study heterogeneity and the The initial search identified a total of 40 studies for ERCC1 C8092A and ERCC2 Lys751Gln, of which 10 studies met the selection criteria Among the 10 selected studies, seven studies reported ERCC1 C8092A, including 2,936 glioma cases and 4,017 controls  [16], , [1P > 0.05). The methodological quality scores of all the included studies were from 12 to 19, indicating that all the included studies have moderate or high quality.The genetic distributions of ERCC1 C8092A and ERCC2 Lys751Gln polymorphisms in the all studies followed the law of HWE (P > 0.1). In addition, no between-study heterogeneity was identified using I2 statistic (P = 0.01). Significant results were also observed in recessive model . There was no significant association in other genetic models. Subgroup analysis showed that ERCC1 8092AA was significantly associated with the risk of glioma compared with ERCC1 8092CC in Chinese population , but was not found in Caucasian population   . Similar = 0.12) .P = 0.03), and large between-study heterogeneity was identified by I2 statistic (P = 0.02), but this was not found in the Chinese population   .Next, stratified analyses for different types of glioma including glioblastoma multiforme and nonglioblastoma multiforme were performed in ERCC2 Lys751Gln polymorphism. There was no significant association in both glioblastoma multiforme and nonglioblastoma multiforme groups .P = 0.02). We then estimated the influence of individual studies on the combined results by omitting one study at a time. No significant differences were found in different genetic models for ERCC1 C8092A polymorphism. When deleting a study in the Chinese population P = 0.04, OR = 0.89, 95%CI: 0.79–0.99). Therefore, our results were generally and statistically reliable.To further confirm the combined results, a sensitivity analysis was conducted by changing the fixed or random effects model. It was shown that the combined results between fixed effects model and random effects model for ERCC1 C8092A polymorphism were no different. However, a significant change was identified in recessive model for ERCC2 Lys751Gln polymorphism. The combined OR (95%CI) was 0.88 (0.79–0.98) estimated by fixed effects model  and ERCC2 Lys751Gln polymorphisms . In the funnel plot . When the analysis was stratified by ethnicity, a significant association was identified in the Chinese population with combined OR of 1.37 (1.03–1.81). However, the significant association disappeared for Caucasians. Similar results were identified in the recessive model of ERCC1 C8092A polymorphism. Remarkably, none of the original researches in Chinese populations reported the significant association between ERCC1 C8092A polymorphism and the risk of glioma. Therefore, the positive results of this meta-analysis might have been caused by small sample size. However, ERCC1 C8092A polymorphism influence the expression of ERCC1 and the DNA repair function of NER pathway may be weakened, a possible explanation of our findings from the point of the pathogenesis For ERCC2 Lys751Gln polymorphism, no significant association between ERCC2 Lys751Gln polymorphism and the risk of glioma was observed. However, TT genotype in ERCC2 Lys751Gln polymorphism may significantly decrease the risk of glioma in the Caucasian population with combined OR of 0.87 (0.78–0.97), but not in the Chinese population. Although 2230 glioma patients and 3393 controls were included in the subgroup analysis in the Caucasian population, only one study with larger sample size presented significant results , which may influence the combined results I2 statistic were used to test the between-study heterogeneity. For all studies, the significant heterogeneity existed only under recessive model in ERCC2 Lys751Gln polymorphism. When the studies were stratified according to ethnicity, significant heterogeneity still existed in the Chinese population, but no statistically significant heterogeneity was observed in the Caucasian population. This suggests that significant heterogeneity might be attributed to differing ERCC2 Lys751Gln polymorphism by ethnicity.In this study, the heterogeneity Q statistic test and the Sensitivity analysis was performed by changing the fixed or random effects model and omitting one study each time to observe the change of effects. For ERCC1 C8092A polymorphism, no significant change was observed. However, the effect of ERCC2 Lys751Gln polymorphism on the risk of glioma changed significantly, though the combined OR was very close to 1. We then looked for publication bias using Begg's test, Egger's test and funnel plot. No significant publication bias existed in this meta-analysis. This indicated that the results in this meta-analysis were generally reliable.Although publication bias was not identified in this meta-analysis, there are some potential limitations. First, only ten eligible studies were included in this meta-analysis. Therefore, in the subgroup analyses by ethnicity, the number of cases and controls was relatively small, which may lead to low statistical power in identifying the association. Furthermore, different pathological types of glioma were not considered, which may be the source of between-study heterogeneity. Unfortunately, only two studies in Caucasian populations provide genetic data of ERCC2 Lys751Gln polymorphism for glioblastoma multiforme and nonglioblastoma multiforme, respectively. Lastly, our meta-analysis was largely performed by unadjusted estimates, because of the limitations in selected studies that presented adjusted estimates. Although adjusted estimates were shown, the estimates were not adjusted by the same confounders. It was also difficult to present the combined estimates by adjusted potential confounders.In conclusion, our meta-analysis suggested that ERCC1 8092AA genotype was associated with the higher susceptibility of glioma in the Chinese population. However, TT genotype of ERCC2 Lys751Gln polymorphism might decrease the risk of glioma in Caucasian population. Due to the small number of studies and moderate methodological quality, the results require cautious interpretation. Studies with larger sample size and more specified information in pathological types of glioma are needed to confirm our results for both Chinese and Caucasian populations.Checklist S1PRISMA checklist.(DOC)Click here for additional data file.


It takes the form of an amino acid substitution, from serine to cysteine, in codon 326. Several epidemiological association studies have been conducted on this polymorphism and its relationship with the risk of prostate cancer. However, results have been conflicting. To resolve this conflict, we conducted a meta-analysis on the association between this polymorphism and prostate cancer, taking into account race, country, sources of controls, and smoking status. A total of nine studies covering 2779 cases and 3484 controls were included in the current meta-analysis. Although no significant association was found between hOGG1 Ser326Cys polymorphism and prostate cancer susceptibility in the pooled analysis, individuals with Ser/Cys+Cys/Cys genotypes were found to have greater risk of prostate cancer if they were also smokers  rather than non-smokers , compared with those with Ser/Ser genotype. In conclusion, our meta-analysis demonstrates that hOGG1 Ser326Cys polymorphism is a risk factor for prostate cancer in smokers. Further studies are needed to confirm this relationship.The HOGG1 gene catalyzes the excision of modified bases and removal of DNA damage adducts. It may play an important role in the prevention of carcinogenesis. Ser Oxidative DNA damage is involved in carcinogenesis. It causes mutations that can inactivate tumor suppressor genes and activate oncogenes. The major form of DNA adduction induced by oxidative damage is 8-OH-dG (8-hydroxy-2-deoxyguanine) and increased 8-OH-dG formation in DNA results in mutagenesis and carcinogenesis Prostate cancer is most commonly-diagnosed malignancy in elderly men in developed countries, and the incidence increases every year. Deficient DNA repair mechanisms may play a role in the age-related increase in prostate cancer risk by allowing carcinogenic DNA damage events to accumulate uncorrected. Higher levels of 8-OH-dG and downregulation of hOGG1 also have been observed in benign prostatic hyperplasia (BPH) and prostate cancer.326Cys, rs1052133). The hOGG1 protein encoded by the Ser326 allele showed much more DNA repair activity than that encoded by Cys326 allele in vitro326 allele had a significantly increased risk of prostate cancer, but Xu et al. found that subjects with Cys326 allele had a reduced risk of prostate cancer. For this reason, we conducted a meta-analysis on hOGG1 Ser326Cys polymorphism and the risk of prostate cancer, taking into account certain characteristics of the subjects and studies, such as race, country, source of controls and smoking status. We believe that this will help us better understand the risk of prostate cancer.Variations in the hOGG1 gene have been identified and the repair activities of the variant proteins have been evaluated in several studies. One of the most frequently analyzed variations is in exon 7 of the hOGG1 gene, which takes the form of a single amino acid substitution, from serine to cysteine at codon 326 (SerThe two online bibliographic databases (PubMed and Embase) were consulted with the following search strategy: “human oxoguanine glycosylase 1, hOGG1 or OGG1, hOGG or OGG,” “polymorphism or variant,” and “prostate cancer or prostate neoplasm” . Original studies in English on hOGG1 polymorphism in prostate cancer were included; reviews, editorials, and letters were excluded. All the references of relevant reviews and eligible articles that our search retrieved were checked carefully.326Cys polymorphism and the risk of prostate cancer. (ii) Each study had to use a case-control design. (iii) Each study had to contain information about available genotype frequency that could help technicians infer the results from the papers. For each of the eligible case-control studies, the following data were collected: the first author's last name, year of publication, country of origin of the subjects, race of the subjects, sources of controls, smoking status of the subjects, number of genotyped cases and controls, and Hardy-Weinberg equilibrium status.Two investigators searched the literature and extracted data independently. The inclusion criteria were as follows: (i) Each study had to discuss or concern hOGG1 Ser2-based Q-test was used to check the heterogeneity of the current study and determine the methods for calculating OR. If P>0.05 for a given Q-test indicated a lack of heterogeneity among the studies, then the summary ORs were calculated using the fixed-effects model (the Mantel-Haenszel method). Otherwise, the random effects model (DerSimonian and Laird method) was used. The significance of the pooled OR was determined using the Z-test, and P<0.05 was considered statistically significant. Subgroup analyses were also conducted on the basis of subject race, country, source of controls, and smoking status. The statistical power was calculated using PS software (http://biostat.mc.vanderbilt.edu/twiki/bin/view/Main/PowerSampleSize).The association between hOGG1 different genotypes (including the heterozygote comparison (Ser/Cys vs. Ser/Ser) and the homozygote comparison (Cys/Cys vs. Ser/Ser), the dominant genetic model (Ser/Cys+Cys/Cys vs. Ser/Ser) and the recessive genetic model (Cys/Cys vs. Ser/Cys+Ser/Ser)) and susceptibility to prostate cancer was measured using the crude odds ratio (OR) with related 95% confidence interval (CI). A χThe publication bias was determined using Egger's linear regression test by visual inspection of the Funnel plot. All statistical analyses were performed using Stata 10.0 .Characteristics of eligible studies are presented in Four studies had been conducted on Caucasians, one on Asians, one study on Africans, and three in mixed populations (Caucasians and African Americans). Six studies were population-based 326Cys polymorphism and the risk of prostate cancer are shown in Pheterogeneity/P = 0.047/0.409), Cys/Cys vs. Ser/Ser , Ser/Cys+Cys/Cys vs. Ser/Ser , or Cys/Cys vs. Ser/Cys+Ser/Ser  in the pooled analysis  rather than non-smokers , relative to those with the Ser/Ser genotype.Genotype information of the Sert = 2.08, P = 0.076 for Ser/Cys vs. Ser/Ser).Sensitivity analysis was conducted to assess the influence of each study on the pooled OR by omission of individual studies. As shown in 326Cys polymorphism as a determinant of prostate cancer risk was investigated in a sample of 6263 subjects from nine published case-control studies. However, no significant association was found under any genetic model in the overall analysis. Ever since the hOGG1 Ser326Cys polymorphism was found to occur frequently in human populations, studies on this polymorphism and the risk of cancer have been conducted and published. Xu et al. first found this polymorphism to be associated with the risk of prostate cancer in a Caucasian population in 2002 326Cys polymorphism and prostate cancer in an African-Caucasian-mixed population. In the current study, we retrieved 9 papers with more subjects and we also stratified the analyses not only by race but also by country, source of controls, and smoking status. We found that the Cys326 allele was associated with a more pronounced increased risk of prostate cancer in smokers than in non-smokers.The potential role of hOGG1 Ser326 allele may be associated with an increased risk and severity of many types of cancer 326 allozyme lacks the ability to repair the DNA damage induced by environmental chemicals in carcinogenesis. However, the exact repair function associated with this polymorphism remains unclear. Kohno et al. observed that the Cys326 allele was less capable of complementing a repair-deficient strain than the Ser326 allele, while Dherin et al. found no significant differences in hOGG1 protein enzymatic activity in vitro 326Cys polymorphism 326 allele was not associated with the risk of prostate cancer, confirming that the enzymatic activity of hOGG1 might not be determined by Ser326Cys polymorphism alone. Rather, it is possible that other confounding factors might interact with this polymorphism in the process of DNA repair.HOGG1 is a key protein involved in base excision repair. It recognizes and excises lesions from oligodeoxynucleotides with DNA damage. Recently, studies have suggested that the hOGG1 Cys326Cys polymorphism, cigarette smoking, and risk of prostate cancer. We found that the risk effect of Ser/Cys+Cys/Cys genotypes was more pronounced among smokers than among non-smokers. The powers of the subtype analyses were 0.60 in the non-smoker group and 0.95 in the smoker group, suggesting an interaction between the hOGG1 polymorphism and smoking with respect to prostate cancer. Such an interaction is reasonable considering that the hOGG1 protein protects cells against the reactive oxygen species and DNA adducts that can be caused by from cigarette smoking. The Cys326 allele has been reported to be associated with decreased DNA repair activity. For this reason this allele was thought to be a risk factor for smokers.Although cigarette smoking has been found to be involved in many cancers and is one of the leading causes of cancer-related death, its role in prostate cancer remains ill-defined The current study has some shortcomings that should be addressed. First, the U.S., Canadian, and Australian populations were classified as Caucasian. However, populations in these areas consist both of indigenous peoples and many types of immigrants, and not all of the studies stated the ancestries of their participants clearly. Second, the controls were not uniform either. In hospital-based studies, most of the controls were BPH patients. Some individuals in the control group might be likely to develop cancer in subsequent years, even if they showed no clinical symptoms at the time of investigation. Misclassification bias can cause deviated genotype distribution in the controls. In spite of these, the present meta-analysis also had some advantages over previous studies. For the first time, we conducted a meta-analysis on hOGG1 polymorphism on prostate cancer susceptibility while considering possible confounding factors, such as smoking status, by polling the results of all published independent studies. The statistical power of the analysis is greater than that of any single study, although it remains relatively small. The quality of studies included in our meta-analysis was satisfactory and perfectly met our inclusion criteria.326Cys polymorphism is a risk factor for prostate cancer in smokers. Further studies are needed to confirm the relationship. We believe that these results will foster comprehensive understanding of the association between hOGG1 polymorphism and the risk of prostate cancer with respect to gene-gene and gene-environment interactions.In conclusion, our meta-analysis demonstrates that hOGG1 Ser


Kaposi's sarcoma-associated herpes virus is associated with primary effusion lymphoma and multicentric Castleman's disease.Seropositivity to lytic and latent Kaposi's sarcoma herpes virus (KSHV) antigens were examined in 2083 lymphomas and 2013 controls from six European countries.P>0.05). The KSHV seropositivity was associated with splenic marginal zone lymphoma (SMZL) (odds ratio (OR)=4.11, 95% confidence interval (CI)=1.57–10.83) and multiple myeloma .Antibodies against KSHV latent and lytic antigens were detectable in 4.5% and 3.4% of controls, respectively, and 3.6% of cases (The KSHV is unlikely to contribute importantly to lymphomagenesis among immunocompetent subjects. However, the observed association with SMZL may underline a chronic antigen mechanism in its aetiology. Kaposi's sarcoma herpes virus (KSHV) also known as human herpes virus-8 is tightly associated with the development of Kaposi sarcoma (KS) and primary effusion lymphoma (PEL), and there is limited evidence of an association with multicentric Castleman's disease (MCD) (Literature is scarce in relation between KSHV infection and the development of non-Hodgkin lymphoma (NHL). In 2004, a case–control study carried out among immunocompetent patients in Spain failed to identify a major contribution of KSHV infection to lymphomagenesis. Although based on few cases, KSHV seropositive patients were more likely to have lymphoplasmacytic lymphoma and low-grade lymphoma (The aim of this study was to investigate the association between KSHV seropositivity and lymphoma subtypes in the multicenter European case–control study EpiLymph.n=2118) of cases and 82% (n=2048) of controls. After excluding immunosupressed patients, 2083 cases and 2013 controls were included in this study.Information was collected on 2362 incident lymphoma cases and 2465 controls during 1998–2003 in six countries . In Germany and Italy, population-based controls were sampled whereas, in all the other countries, hospital-based controls were recruited. The participation rate was 68.5% and 87.7% among controls and cases, respectively. The study has been described elsewhere  and 95% confidence interval (95% CI) of lymphoma in relation to KSHV seropositivity. All models were adjusted for age, sex and country. Two-sided P-values were considered statistically significant at the 0.05 level. Heterogeneity among countries was evaluated with a likelihood ratio test comparing the model with and without interaction between country and exposure. We used the kappa (κ) statistic to describe the agreement between assays and the results of the two assays.Differences in KSHV seropositivity by categorical variables were done by means of a We carried out sensitivity analyses to evaluate the influence of country, type of control, cutoffs for seropositivity and age on our results, and to reduce the possibility of misclassification of the KSHV status. All the analyses were conducted using STATA  and R .P=0.5), increased linearly with age (P-trend <0.0001) and decreased with increasing educational level (P-trend=0.03) for latent antibodies. The highest seroprevalence was observed in Italy (12%). Similar pattern for K8.1 antibodies was observed in relation to demographic characteristics. Seropositivity among controls for both antibodies represented 2% (n=38). Overall, the pairwise agreement between ORF73 and K8.1 results among controls was moderate (κ=0.46), being good in the youngest (κ=0.61) and in the age group of 64 to 70 years old (κ=0.63) and in Italy (κ=0.74) (data not shown).N=75/2083) nor to K8.1 . By lymphoma subtype, CLL/SLL and MM cases were more likely to be seronegative to latent anti-KSHV antibodies than controls . Antibodies against K8.1 antigen, but not for ORF73 antigen, showed a significant increased OR for splenic marginal zone lymphoma (SMZL) . Although 70% (N=25) of SMZL cases came from Spain, sensitivity analyses on country did not modify the results (data not shown). We also observed a nonsignificant increased OR for mantle cell lymphoma (MCL) . Further sensitivity analyses on age, type of control and OD cutoff variations did not change the results.This study analysed the association between antibodies against KSHV and lymphoma in the European case–control study EpiLymph. Overall, we found no association between positivity to lytic or latent antibodies against KSHV antigens and lymphoma. However, we observed an increased OR in SMZL and MCL, and a decreased OR in MM. Furthermore, our data support previous observations in which seropositivity to KSHV increases with increasing age and is higher in Italy than in other European countries (We observed that KSHV seroreactivity to lytic antigens increased four times the probability of SMZL. This was consistent with our previous results based on Spanish cases (In our data, we detected a nonsignificant increased OR in MCL. To our knowledge, this has not been previously reported and we do not have a clear biological explanation. The MCL may be subject to histological classification . HoweverThe observed reverse association of KSHV and MM is consistent with the literature (To our knowledge, this is the first study that evaluates the association between anti-KSHV antibody status and lymphoma among immunocompetent European subjects. We have enlarged the sample size of our previous study, including participants from four European countries to validate our previous finding and also to explore further the association with specific lymphoma subtypes. Sensitivity analyses, including variation of seropositivity cutoffs and age-matched analyses, supported that our results were not materially influenced by potential confounding factors. Serological evaluation of KSHV has not been standardised (In conclusion, our findings support an association with SMZL among immunocompetent subjects that deserves further research.


Cathepsin D C224T polymorphism has been reported to associate with AD susceptibility. But the results were inconsistent. This study aimed to assess the relationship between C224T polymorphism and AD risk.The relevant studies were identified by searching PubMed, Embase, Web of Science, Google Scholar and Wan fang electronic databases updated on July 2013. The relationship between Cathepsin D C224T polymorphism and AD risk was evaluated by ORs and 95% CIs.A total of 25 case-control studies including 5,602 cases and 11,049 controls were included in the meta-analysis. There was no association between C224T polymorphism and AD risk with all the studies were pooled in the meta-analysis . Furthermore, when stratified by ethnicity, age of onset and APOEϵ4 status, significant association did not found in all subgroups.The present meta-analysis suggested that the Cathepsin D C224T polymorphism was not associated with AD susceptibility. APOEϵ4) was the only verified risk factor for sporadic AD [APOEϵ4 allele was neither necessary nor sufficient for AD development. About 50% of AD patients did not have mutations in the genes mentioned above or carry the APOEϵ4 allele, and not everyone who has the mutations of the genes will acquire AD [The neurodegenerative disorder Alzheimer’s disease (AD) caused the most of dementia in the elderly . Previouradic AD . Howeverquire AD , suggestCTSD), an intracellular acid protease, contributed to the proteolytic cleavage of APP and the clearance of the β-amyloid (Aβ) from the central nervous system [CTSD might involve in the pathogenesis of AD. Variants of CTSD gene might impede the functions of proteolytic degradation, thus increasing the risk of AD. A CTSD C224T polymorphism(C-to-T) in exon 2 can bring about amino acid change , increase pro-CTSD secretion and alter intracellular maturation [The main histopathologic features of AD are neurofibrillary tangles and Neuritic plaques which consist of hyperphosphorylated tau protein and amyloid peptides, respectively. Cathepsin D inclusion of the sample size and distribution of alleles and genotypes; (4) AD diagnosed according to the criteria of the National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer’s Disease and Related Disorders Association (NINCDS-ADRDA), or the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV). Exclusion criteria of our study were followed as: (1) duplicated literature, reviews, or animal studies; (2) genotype frequency and distribution were not included; (3) not enough information for data extraction.The eligible studies were requested to agree with the inclusion criteria: (1) a case–control study; (2) research of the correlation between Two reviewers (Cuiju Mo and Jingzhe Sui) extracted the information independently. If there was a disagreement, the data was checked again, and a third reviewer (Xue Qin) was invited to check the data. Information collected from each eligible study was included: first author, year of publication, country, ethnicity, genotyping method, AD diagnosis, control sources, sample sizes, age of onset, and genotype distribution in cases and controls.CTSD with the APOEϵ4 allele, we compared the dominant genetic model (TT + TC vs. CC) between case and control subjects stratified by the APOEϵ4 allele status. Similarly, the relationship of the APOEϵ4 allele with AD risk was investigated between the patients carrying the T allele or not.All analysis was conducted using Stata version 12.0 software . The association was assessed by pooled odds ratio (OR) together with the corresponding 95% confidence interval (CI). Only heterozygote comparison model (TC vs. CC) and the dominant genetic model (TT + TC vs. CC) were analysed. Furthermore, we evaluated the effect in different subgroup stratified by ethnicity (Asian vs. Caucasian) and age of onset. Early-onset AD (EOAD) was defined as age at onset <65 years, and age at onset ≥65 years was considered as late-onset AD (LOAD). To evaluate the interaction of the 2-test based Q-statistic and I2 statistic was used to evaluate the heterogeneity among the studies. The DerSimonian–Laird random-effects model was used to assess pooled OR when a significant heterogeneity (PQ < 0.1 or I2 ≥ 50%) was observed. Otherwise, the Mantel–Haenszel fixed-effects model was used. The publication bias was detected by funnel plot and Egger’s test. An Egger’s test P value <0.05 was considered as statistically significant. The genotype distribution of the control population was used to evaluate Hardy–Weinberg Equilibrium (HWE) by a goodness-of-fit Chi-square test. P <0.05 (two-side) was considered as statistically significant.The xCTSD and the APOEϵ4 allele [Figure 4 allele ,28,29,344 allele ,34. The C224T polymorphism was not associated with AD risk. The heterogeneities of CT vs. CC and the dominant CT + TT vs. CC models were assessed in the overall population, and the PQ values were 0.023 and 0.007, respectively. Thus, random-effects model was chose to analyse the CT vs. CC model  and Caucasian. Similarly, we found non-significant associations in the EOAD  and LOAD subgroups in any of the comparisons .There was no visible publication bias among the studies because of the shape of the Begg’s funnel plots revealed symmetry in the CT vs. CC and CT + TT vs. CC comparative genetic models Figure . StatistCTSD polymorphism and AD risk from abundant data over 16,651 genotype cases and controls.The effects of genetic sequence variants in complex human traits are not readily detectable in population samples. However, meta-analysis that accumulates published data from small single research is a valuable tool in identifying disease genes. The functions of CTSD are to hydrolyse APP protein and clear Aβ from the central nervous system ,11. In ACTSD C224T polymorphism and AD susceptibility. Our finding indicated that the C224T polymorphism was not associated with the AD risk both in Asian and Caucasian populations, which were in accord with the results of the previous meta-analysis [TT polymorphism, and the proportion of TT was very small, as is usual in common polymorphisms, heterozygote might be responsible for the significant difference in frequency; therefore, we only compared the CT vs. CC and the dominant CT + TT vs. CC models. Lastly, no significant publication bias was observed in any of the studies analyzing by Egger’s test and Begg’s funnel plot. Thus, based on the above factors, the results of our meta-analysis were more reliable than those of previous studies.As far as we know, the present meta-analysis involving 5602 cases and 11,049 healthy controls was the most comprehensive to date to investigate the relation between the analysis  and incoanalysis . CompareCTSD C224T polymorphism and AD risk. Given that the control genotypes of two case-control studies [2 = 39.65, PQ = 0.023) and CT + TT vs. CC  in the overall populations. Several factors might contribute to the heterogeneity. First, AD is a complicated and multi-genetic disease. Second, clinical heterogeneity, such as gender, age of onset, and diagnosis criteria, were factors. The different studied populations, such as ethnicity, might also explain the discrepancy. In subgroup analysis stratified by ethnicity and age of onset, heterogeneity only existed in the Caucasian subgroup, indicating that age was the major contributor to the existence of all heterogeneity.Our results from the CT vs. CC and dominant CT + TT vs. CC comparison models suggested that no significant correlation was existed between the  studies  were outCTSD C224T polymorphism and AD risk either in Asian or in Caucasian population, which was inconsistent with the previous meta-analysis[CTSD C224T polymorphism increased AD risk in men only which might cause the false-negative result. As no study has clarified gender-specific differences regarding lysosomes or its components and the characteristic lesions in AD, therefore, future study with larger samples to investigate the gender-specific is necessary. When stratified by age of onset, we found no significant differences both in EOAD and LOAD subsets. Possible explanations for these findings might be the small sample sizes for analysis; the same control source, without strict age matching, and missing age information in some studies. Given these factors may affect the statistical power. Further research is required to assess the gene effects and validate our findings.Considering the impact on the summary OR of different ethnicities, we further performed subgroup analysis based on ethnicity. Those results indicated no significant association between the -analysis. Similar-analysis were exc-analysis and Mate-analysis study, a-analysis study. TCTSD polymorphism and APOEϵ4 allele on AD, ten studies which provided genotype distribution data of APOEϵ4 status were chosen for further study, and of which only four showed evidence of an association [C224T polymorphism and AD risk in APOEϵ4 carriers and non-carriers. The association of CTSD T allele with AD risk between APOEϵ4 carriers and non-carriers in Caucasians was quite similar, contrary to the Schuur result. Due to the lack of an Asian population in the Schuur study, sample size and ethnicity might have contributed to some bias in the final result. While the ORs of APOEϵ4 were greater in the T allele carriers group than the subjects without the T allele. Because of the extensive overlap in two effect sizes and the remarkably small group of subjects who carry both the APOEϵ4 and CTSD T alleles, the association between the CTSD T and APOEϵ4 alleles should be interpreted cautiously.To evaluate the interaction of ociation ,15,20,25C224T polymorphism, discounted the potential linkage disequilibrium with another mutation of this gene, and ignored the interactions between gene and gene or gene and environment. Third, the data of our meta-analysis was unadjusted; the suspected factors could be analysed, such as, gender, diet, lifestyle habit, and environmental factors. Fourth, we included the English or Chinese publications only; the lack of unpublished data and data published in other languages might contribute some bias. There were only four articles in the Asian subgroup, with small sample size, which may cause low statistical power.There were some limitations that merit attention. First, some of the eligible studies lacked sufficient information for detailed and deep analysis. In some studies, the controls were not uniformly defined as matched by age and gender; and it may lead to some negative correlation. Second, we mainly focused on the CTSD C224T polymorphism was not associated with AD risk both in the overall populations and the subgroups stratified by ethnicity and age of onset. In addition, we found no statistically significant differences between the CTSD C224T genotypes and AD stratified by APOEϵ4 allele status. Our data did not suggest that the CTSD C224T polymorphism was a possible susceptibility factor for AD. Future studies will require much larger sample sizes and will need to analyse the impact of this polymorphism in other populations.The finding of our present study revealed that the AD: Alzheimer’s disease; CTSD: Cathepsin D; APOE: Apollipoprotein E; EOAD: Early-onset AD; LOAD: Late-onset AD.All authors state no conflict of interest.CM, QL, XQ, SL originally designed the study, collected the literature data and drafted the manuscript. JS, JW, YD performed data extraction and statistical analysis. LX, TL, YH executed literature search and checked the results. All authors have read and agreed with the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2377/14/13/prepub


MMP) polymorphisms and bladder cancer risk has become a hot topic and was studied extensively in recent years, but the results are still controversial. In order to estimate the relationship of MMP polymorphisms and the risk of bladder cancer, we performed this meta-analysis. We conducted a comprehensive search of databases; PubMed, Web of Science, Embase, Chinese Biomedical Literature Database  and Wanfang Database (Chinese) were searched for all case–control studies which mainly study the relationship between MMP-1-1607 1G/2G, MMP-2-1306 C/T, and MMP-9-1562 C/T polymorphisms and the susceptibility of bladder cancer. The association between the MMP polymorphisms and bladder cancer risk was conducted by odds ratios (ORs) and 95 % confidence intervals (95 % CIs). At last, totally five literatures with 1,141 cases and 1,069 controls were contained in the meta-analysis. Among these articles, four articles with 1,103 cases and 1,053 controls were about MMP-1-1607 1G/2G polymorphism and three studies with 839 cases and 775 controls for MMP-2-1306 C/T polymorphism and MMP-9-1562 C/T polymorphism. With regard to MMP-1-1607 1G/2G polymorphism, significant association was found with bladder cancer susceptibility only under recessive model , and as to the MMP-2-1306 C/T polymorphism, significant association was found with bladder cancer susceptibility only under homozygote model , but no associations was found between MMP-9-1562 C/T polymorphism and bladder cancer susceptibility. The results suggest that the MMP-2-1306 C/T and MMP-9-1562 C/T polymorphisms are significantly associated with bladder cancer susceptibility, and no associations were found between MMP-9-1562 C/T polymorphism and bladder cancer susceptibility.The relationship between matrix metalloproteinase ( Bladder cancer is one of the most common malignant diseases around the world that has the highest recurrence rate of any malignancy –3. The mMMP) enzymes may contribute to the development of bladder cancer [MMP is a family of matrix metalloproteinases and a zinc-dependent endopeptidase enzyme that can reduce substantially all of the extracellular matrix components such as basement membrane, collagen, and fibronectin [MMPs play an important role not only in physiological but also in pathological conditions, including tissue regeneration, wound repair, reproduction, arthritis, atherosclerosis, and so on [MMP-1, MMP-2, and MMP-9 genes are three important members of the MMP family. The polymorphisms of MMP-1-1607 1G/2G, MMP-2-1306 C/T, and MMP-9-1562 C/T were reported to be related with bladder cancer susceptibility, but the conclusions remain to be inconsistent [Previous studies have reported that genetic variants in genes encoding proteins like matrix metalloproteinase ,” and “variation” for all study publications before July 2013, and there was no language restriction in the literature search. Additional studies were identified by a hand search of the references of original research, and reviews were also examined in order to find more eligible studies. With regard to published studies of the same author, with overlapping data, we selected the most recent or complete study only.We conducted a comprehensive search strategy through searching the electronic databases, such as the PubMed, Web of Science, Embase, Chinese Biomedical Literature Database , and Wanfang Database (Chinese) using the following search terms: MMP-1-1607 1G/2G, MMP-2-1306 C/T, and MMP-9-1562 C/T polymorphism and bladder cancer risk; (b) case–control study; (c) provided sufficient genotype data in order to calculate the odds ratio (OR) with 95 % confidence interval; (d) genotype frequencies in controls was abided by Hardy–Weinberg equilibrium (HWE). The following were exclusion criteria: (a) not a case–control study, (b) reviews or case reports, (c) no available data reported, and (d) duplicated reports.The inclusion criteria of our meta-analysis were as follows: (a) evaluation of the Two of our authors independently extract the information from each research report according to the inclusion criteria listed above. When they have different opinions, they would reach agreement through discussion. If they cannot reach consensus, the third author would consulted to resolve the contradiction. The following information were extracted: (a) first author's name, (b) publication years, (c) country of origin, (d) ethnicity, (e) genotyping methods, (f) source of the control group, (g) sample size of cases and controls, and (h) type of genotype. We also evaluated whether the genotype distributions followed the Hardy–Weinberg equilibrium.MMP-1-1607 1G/2G polymorphism with the risk of bladder cancer was evaluated by OR and 95 % confidence interval (CI) according to allele contrast (2G vs. 1G), homozygote (2G2G vs. 1G1G), heterozygote (1G2G vs. 1G1G), recessive (2G2G vs. 1G2G/1G1G), and dominant (2G2G/1G2G vs. 1G1G) models, while the strength of association between the MMP-2-1306 C/T and MMP-9-1562 C/T polymorphisms and bladder cancer susceptibility was evaluated by OR and 95 % CI according to allele contrast (T vs. C), homozygote (TT vs. CC), heterozygote (TC vs. CC), recessive (TT vs. TC/CC), and dominant (TT/TC vs. CC) models, respectively. The heterogeneity was tested by a chi-square-based Q statistic test. The effect of heterogeneity was quantified by using I2 value as well as P value [I2 value is >50 % or P < 0.10, suggesting that an obvious heterogeneity existed, ORs were pooled by random effect model [The possible association between the  P value . If I2 ct model . Otherwict model .http://ihg2.helmholtz-muenchen.de/cgibin/hw/hwa1.pl), if P > 0.05 suggests that the controls followed HWE balance. A sensitivity analysis was carried out to assess whether there is stability of our results. By way of deleting a single study, there was a time to assess whether the study results have an impact on the overall pooled ORs. [P < 0.05 indicates that statistically significant publication bias existed) [We assessed the HWE for the control group in every article by using the professional web-based program (led ORs. . Publicaexisted)  and visuexisted) . All staMMP-1-1607 1G/2G polymorphism, three studies with 839 cases and 775 controls for MMP-2-1306 C/T polymorphism, and three studies with 839 cases and 775 controls for MMP-9-1562 C/T polymorphism; the general characteristics of studies included in the meta-analysis were listed in Table A total of five articles , 19–23 wMMP-1-1607 1G/2G polymorphism, significant association was found with bladder cancer risk only under recessive model  in Fig. The main results of this meta-analysis and the heterogeneity assessment were shown in Tables To estimate the sensitivity of our meta-analysis, a leave-one-out sensitivity analysis was performed. A single article included in the meta-analysis was assessed each time to reflect the impact of the individual data set to pooled ORs. Any single study was omitted, while the overall statistical significance does not change, indicating that the results are stable (data not shown). Therefore, the sensitivity analysis results show that our meta-analysis data is relatively stable and credible.P > 0.05, data not shown) suggest that no publication bias existed in our meta-analysis.Both funnel plot and Egger’s test were conducted to access the publication bias. Funnel plot is relatively straightforward to observe whether the publication bias is presence, and Egger’s test was used to provide a statistical evidence of funnel symmetry. Both the shapes of the funnel plot Fig.  and EggeMMP polymorphisms and bladder cancer susceptibility; however, the results obtained were inconsistent or controversial [Bladder cancer is one of the most common urinary malignant diseases around the world . Howeveroversial , 19–23.MMP-1-1607 1G/2G, MMP-2-1306 C/T, and MMP-9-1562 C/T polymorphisms and bladder cancer risk was explored. Our results suggest that MMP-1-1607 1G/2G polymorphism was significantly associated with bladder cancer risk under recessive model, and significant association was found between MMP-2-1306 C/T polymorphism and bladder cancer risk under homozygote model, but there is no association found between MMP-9-1562 C/T polymorphism and bladder cancer risk. The results indicate that potentially functional MMP-1-1607 1G/2G and MMP-2-1306 C/T polymorphisms may play an important role in the development of bladder cancer.Meta-analysis is a powerful tool that combines the world’s research literature that can resolve the statistical power and discrepancy problem in associated studies . It is aMMP polymorphisms and bladder cancer risk, there are still some limitations that should be pointed out. Firstly, the sample size of each study and the quantity of studies included in our meta-analysis were relatively small. The total sample size may not be enough to make a convincing conclusion. Secondly, there is no uniform definition of the control group. Some studies used population-based controls, while others used hospital-based controls, which may not be a representative of the general population. Thirdly, only published studies were included in our meta-analysis, which is likely to miss some relevant unpublished but may meet the inclusion criteria articles. Thus, publication bias may be incurred. Fourthly, the possible interactions of gene–environment and gene–gene were not assessed in the meta-analysis.In spite of comprehensive analysis conducted to show the association between Therefore, larger-scale and well-designed studies are necessary to estimate the association between MMP polymorphisms and the risk of bladder cancer in the future. Besides, the possible gene–environment and gene–gene interactions should be considered in the future studies too.MMP-9-1562 C/T polymorphism is not associated with bladder cancer. Considering the limitations listed above, larger-scale and well-designed studies are require to further estimate the association between MMP polymorphisms and bladder cancer risk in future studies. Besides, the possible gene–environment and gene–gene interactions should also be considered in future meta-analysis.In conclusion, the results of our meta-analysis suggest that a significant association was found between the MMP-2-1306 C/T and MMP-9-1562 C/T polymorphisms and the risk of bladder cancer, while the


CD14 gene at position -260  C>T has been inconsistently associated with asthma. The aim of this study was to estimate the combined likelihood of developing asthma given the CD14 -260C>T genotype.Asthma is a phenotypically diverse disease with genetic susceptibility. A single nucleotide polymorphism (SNP) in the Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, a systematic search and meta-analysis of the literature was conducted to estimate the association between this SNP and asthma. Planned subgroup analyses were performed to detect potential sources of heterogeneity from selected study characteristics. Post-hoc sensitivity analysis was performed to identify studies exerting excessive influence on among-study heterogeneity and combined effects.Meta-analysis of 23 studies yielded a non-significant overall association with high heterogeneity across studies. After restricting analysis to studies using atopic asthma and non-atopic non-asthma case-control phenotypes and excluding studies influencing heterogeneity, the genotype-specific odds ratios (ORs) suggested a codominant model. Carriers of the TT and CT genotypes were about 33% less likely  and about 20% less likely , respectively, to have atopic asthma compared to carriers of the CC genotype. Among-study heterogeneity may be explained by overly broad asthma phenotype definitions, gene-environment interactions, and gene-gene interactions.CD14 -260T allele and atopic asthma susceptibility was observed. These results demonstrate the importance of precisely specified case-control groups as well as the need to assess interactions in the investigation of complex diseases such as asthma.A protective dose-response relationship between the  CD14) )) was plotted against the OR of each study.Click here for fileCD14 -260 (-159) CT versus CC genotypes for all reviewed studiesFigure S2. Funnel plot of . Standard error of the logarithm of the odds ratio (SE(log[OR])) was plotted against the OR of each study.Click here for fileCD14 -260 (-159) TT versus CT genotypes for all reviewed studiesFigure S3. Funnel plot of . Standard error of the logarithm of the odds ratio (SE(log[OR])) was plotted against the OR of each study.Click here for fileCD14 -260 (-159) TT versus CC genotypes for studies with precisely defined phenotypesFigure S4. Funnel plot of . The funnel plot displays studies included in the review that used atopic asthma cases and non-atopic non-asthmatic controls, excluding heterogeneous studies identified by sequential analysis [analysis ,27,42. SClick here for fileCD14 -260 (-159) CT versus CC genotypes for studies with precisely defined phenotypesFigure S5. Funnel plot of . The funnel plot displays studies included in the review that used atopic asthma cases and non-atopic non-asthmatic controls, excluding heterogeneous studies identified by sequential analysis [analysis ,27,42. SClick here for file


CASR) is associated with the total serum calcium levels. In this study, we collected DNA samples from 480 Taiwanese subjects  for genotyping the CASR gene. Our results indicated no significant association between the CASR polymorphism (rs17251221) and the susceptibility of calcium nephrolithiasis. However, we found a significant association between rs17251221 and stone multiplicity. The risk of stone multiplicity was higher in patients with the GG+GA genotype than in those with the AA genotype . In conclusion, our results provide evidence supporting the genetic effects of CASR on the pathogenesis of calcium nephrolithiasis.Calcium nephrolithiasis is one of the most common causes of renal stones. While the prevalence of this disease has increased steadily over the last 3 decades, its pathogenesis is still unclear. Previous studies have indicated that a genetic polymorphism (rs17251221) in the calcium-sensing receptor gene ( CASR), vitamin D receptor, and osteopontin genes are highly associated with stone formation Nephrolithiasis is a worldwide health problem that affects almost all populations. In Taiwan, its prevalence is approximately 9.6%  CASR causes familial hypocalciuric hypercalcemia (FHH), which leads to life-long hypercalcemia CASR genetic polymorphism (rs17251221) is strongly associated with serum calcium concentration in individuals of European and Indian-Asian descent CASR (rs17251221) on susceptibility to nephrolithiasis is still unclear.Hypercalciuria is one of the main risk factors for nephrolithiasis. Most patients with nephrolithiasis have calcium-containing stones CASR polymorphism (rs17251221) is associated with the development of calcium nephrolithiasis. The number of renal stones is related to the disease activity of nephrolithiasis; therefore, we investigated the association between the CASR genetic polymorphism and the number of renal stones in patients with nephrolithiasis.The aim of the present study was to investigate whether the We conducted a case–control study from February 2009 to December 2010 in Kaohsiung Medical University Hospital, Kaohsiung, Taiwan. In total, 189 patients with calcium nephrolithiasis and 291 controls were included in the study. Calcium nephrolithiasis was diagnosed on the basis of ultrasonographic and radiographic findings. A 20-mL whole blood sample and a one-spot midstream urine sample from the first voided urine were collected from all subjects after they had fasted overnight. Blood and urine samples were used to measure total calcium, phosphate, uric acid, and creatinine levels. All urinary parameters were corrected by using urinary creatinine levels. If stone specimens were removed by surgery or obtained after medical treatment or shock-wave lithotripsy, infrared spectroscopy  was used to confirm the composition of calcium-containing stones Genomic DNA was extracted from whole blood samples by using a standard protocol. Whole blood samples from patients and controls were centrifuged at 3000 rpm for 10 min at 4°C. Buffy coat was isolated from the blood samples. After lysing the red blood cells in a lysis buffer, the samples were mixed with a cell lysis buffer for several days. Protein precipitation solution was added to precipitate the proteins. Finally, total genomic DNA was isolated by precipitation with 95% isopropanol and 80% alcohol.CASR (rs17251221) was selected on the basis of research findings reported by O'Seaghdha et alA single nucleotide polymorphism (SNP) in P>0.05) for both patients and controls. The statistical differences between the patient and control groups or between the multiple and single stone-episode groups in genotype and allele frequency were analyzed by the chi-square test. Under certain circumstances, Yates' correction for the chi-square test was also applied for a more stringent analysis. The relationship between kidney stone diseases and multiplicity and CASR polymorphisms was assessed by the odds ratio (OR). Quantitative variables were expressed as the mean ± standard deviation. The t-test was used to compare the differences between the means of continuous variables. A P value < 0.05 was considered statistically significant. Statistical analyses were performed using SPSS, version 14.0 .The distribution of genotypes was in Hardy–Weinberg equilibrium  associated with the risk of stone multiplicity in calcium nephrolithiasis.Nephrolithiasis is one of the most common urologic diseases in the developed countries. Chemical composition analysis of urinary stones has indicated that calcium oxalate stones are the most common and occur in approximately 75% to 80% of cases CASR is strongly associated with total serum calcium levels CASR variants modulate serum calcium levels in the adult general population CASR genotype on clinical biochemical parameters we did not use a control group. The availability and use of data from the controls would have strengthened our observation. Another possible explanation for our results is the different genetic backgrounds of the populations and the phenotypic heterogeneity of nephrolithiasis. Our findings still need to be replicated in a larger sample size.Nephrolithiasis research has focused on biochemical or metabolic alterations in local urinary constituents leading to stone formation. However, abnormalities in the chemical composition of urine alone do not explain many aspects of nephrolithiasis urinary stone disease CASR gene mutations have been implicated in several rare familial genetic disorders. For instance, the gain-of-function mutations E128A  and T151M  cause an autosomal dominant form of hypocalcemia (MIM #146200), whereas R990G  is a gain-of-function mutation that was reported to predispose patients to primary hypercalciuria and kidney stone disease CASR and subsequent hypercalcemia and hypermagnesemia et al have indicated that the minor G allele of rs17251221 was associated with higher serum calcium and magnesium levels and suggested that the causal G variant may underlie a reduction in CASR activity et al pointed out that there was another coding SNP in this gene, rs1801725 (A986S), which has been detected among families with FHH and in the general population In conclusion, our data showed a significant association between rs17251221 and stone multiplicity in nephrolithiasis patients, suggesting that rs17251221 is as a susceptibility marker for stone multiplicity in nephrolithiasis. Moreover, this is the first evidence that the risk of developing renal stones was greater in nephrolithiasis patients having the G allele of rs17251221 than those having the A allele.


Aim. To study the impact of birth characteristics on the risk of atopic dermatitis in a twin population. Methods. In a population-based questionnaire study of 10,809 twins, 3–9 years of age, from the Danish Twin Registry, we identified 907 twin pairs discordant for parent-reported atopic dermatitis. We cross-linked with data from the Danish National Birth Registry and performed cotwin control analysis in order to test the impact of birth characteristics on the risk of atopic dermatitis. Results. Apgar score, OR (per unit) = 1.23 (1.06–1.44), P = 0.008, and female sex, OR = 1.31 (1.06–1.61), P = 0.012, were risk factors for atopic dermatitis in cotwin control analysis, whereas birth anthropometric factors were not significantly related to disease development. Risk estimates in monozygotic and dizygotic twins were not significantly different for the identified risk factors. Conclusions. In this population-based cotwin control study, high Apgar score was a risk factor for atopic dermatitis. This novel finding must be confirmed in subsequent studies. Atopic dermatitis is a chronic relapsing skin disease with a lifetime prevalence of around 20%. The disease has an early onset with 60% being affected during the first year of life and 85% before the age of five . Atopic The onset of atopic dermatitis early in life implies that risk factors for the disease exert their effect already in utero or in very early childhood. However, previous studies of the association between perinatal factors and the risk of atopic dermatitis are conflicting. With regard to alcohol intake during pregnancy, results indicate an increased risk for atopic dermatitis in children of mothers who consume alcohol during pregnancy , 8. ContThe cotwin control study design is a matched case-control study that utilizes twin pairs discordant for an exposure or an outcome. Particularly, because twin pairs are inherently matched on several confounding factors relating to early life and upbringing, this type of study design is ideal to tease out causative relationships in complex diseases. In the present study we use the cotwin control design to identify birth-associated risk factors for atopic dermatitis in twin pairs discordant for the disease.The study population comprised all live-born twins in Denmark between 1994 and 2000 included in the Danish Twin Registry. In 2003 when the twins were 3–9 years of age the twins' parents responded to a two-page questionnaire concerning atopic diseases, diabetes, febrile and epileptic seizures, and mental health attributes. Atopic dermatitis was diagnosed by the question “has your child ever had eczema in the folds of the elbows or knees?” Zygosity was determined by information regarding the twins' similarity to family, friends, teachers, and relations. This method determines zygosity correctly in more than 95% of the cases compared with genetic marker information . We crosA total of 10,809 twin subjects participated in the questionnaire study (participation rate 68%). Of these, complete information on atopic dermatitis was available for 5399 intact twin pairs: 850 monozygotic (MZ) twin pairs, 2,279 dizygotic (DZ) twin pairs of the same sex, 1,999 DZ twin pairs of opposite sex, and 271 twin pairs of unknown zygosity. There were 907 twin pairs discordant for atopic dermatitis, and of these 58 and 827 were monozygotic (MZ) and dizygotic (DZ) pairs, respectively. t-tests and chi-square tests, ignoring the familial relationship between the twins. Next, the risk factors that were unshared between the twins within the pair were examined in a 1 : 1 matched conditional logistic regression analysis  in order to determine the effect of these risk factors on atopic dermatitis. The factors entered into this analysis were birth weight, birth length, Apgar score, and sex. The matching was done with the affected (atopic dermatitis) twin in each pair being the case and the unaffected twin being the control. DZ same-sex and DZ opposite-sex twin pairs were considered together in the analysis. The matching of the twins in the cotwin control design allows an inherent adjustment for other factors that would otherwise confound the association between birth factors and atopic dermatitis such as genetic factors, atopic predisposition, maternal smoking during pregnancy, gestational age, mode of delivery, and current age, as well as effects of the rearing environment. Therefore the estimated risk of atopic dermatitis can be assumed to reflect more precisely the influence of birth factors, for example, Apgar score. The following predictions can be made for the cotwin control analysis , nonsmoking of the mother during pregnancy (P = 0.002), and monozygosity (0.012) were also significant predictors for atopic dermatitis in the whole sample when ignoring the familial relationship between the twins , P = 0.012, were significantly predisposing factors for atopic dermatitis, whereas birth anthropometric factors were not significantly related to development of atopic dermatitis. Risk estimates in discordant MZ and DZ twins were not significantly different . We had complete ascertainment of all live-born twins in Denmark between 1993 and 2000 and cross-linked these data to data from the Danish Birth Registry combining information from two independent sources. Data on perinatal risk factors were obtained by professional midwifes' records, which increased the validity of the data. Contrary to this, the diagnosis of atopic dermatitis was self-reported by the parents of the twins, which could have induced recall and reporting bias. Moreover, the question used to diagnose atopic dermatitis has not been systematically validated. Another limitation alludes to the differing age at ascertainment of the eczema diagnosis, which may have influenced the results. However, age was automatically eliminated as possible confounder in the cotwin control analysis model since it was the same for the two twins within the pair. The fact that we studied twins may have had implications for the generalisability of the results to the population as a whole. Particularly, twins differ from singletons in a number of ways related to prenatal and early life that potentially could affect the risk of atopic dermatitis differentially in twins. These factors, which may be related to gestation, intrauterine environment, and mode of delivery, could also be expected to influence the variation in Apgar score. However, in a matched case-control study of 112 infant twins and singletons from the United States 5-minute Apgar score was not different in twins compared with singletons when matched for gestational age and mode of delivery , suggestWe conclude that a high Apgar score may be linked to development of atopic dermatitis. Although this finding agrees with the theory that robust babies of high SES are at increased risk of atopic dermatitis, this has not been previously documented and awaits support from future studies.


TRAF5 and TRAF3IP2 have been reported to be associated with several autoimmune diseases. Behçet's disease (BD) and Vogt-Koyanagi-Harada (VKH) syndrome are two autoimmune uveitis entities whereby both genetic and environmental factors are thought to be involved.The role of TRAF5 and TRAF3IP2 in BD and VKH has not yet been reported and was therefore the subject of this study.The study included 789 BD patients, 940 VKH patients and 1601 healthy unrelated individuals. Genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) or TaqMan® SNP Genotyping Assay. Real-Time PCR was used to detect mRNA expression from PBMCs obtained from healthy controls with (n = 22) or without (n = 79) stimulation. Levels of TNF-α, IL-6 and IL-8 in culture supernatants were measured by ELISA (n = 22).−12 to 0.027). TRAF3IP2 rs33980500 and rs13190932 were not polymorphic in Han Chinese. Following stimulation by lipopolysaccharide (LPS), carriers of the GG genotype of rs6540679/TRAF5 had a higher TRAF5 mRNA expression (p = 0.004) and an increased TNF-α (p = 0.0052) and IL-6 (p = 0.0014) level compared with AA and AG genotype carriers.Three SNPs  of TRAF5 and rs13210247 of TRAF3IP2 were significantly associated with Behçet's disease and VKH syndrome (corrected P values ranging from 9.45×10This study provides evidence that TRAF5 and TRAF3IP2 genes are involved in the development of BD and VKH syndrome. Functional research suggested that TRAF5 gene polymorphisms may regulate TRAF5 expression and downstream inflammatory cytokines such as TNF-α and IL-6. Uveitis is an intraocular inflammation that is caused by either infectious or noninfectious mechanisms and which can lead to serious visual impairment. To unravel the immunological mechanisms that cause uveitis we have focused on two commonly occurring uveitis entities in China, namely Behcet's disease (BD) and Vogt Koyanagi Harada (VKH) syndrome BD is an autoinflammatory disease characterized by a diverse spectrum of clinical manifestations including recurrent oral aphthae, uveitis, multiform skin lesions and genital ulceration VKH syndrome is a multisystem autoimmune disorder directed against melanocytes and is characterized by bilateral granulomatous uveitis with meningeal irritation sign (MIS), dysacusis, vitiligo, poliosis and alopecia In our search for a possible role of as yet not identified polymorphisms in immunoregulatory genes in the pathogenesis of uveitis, we assessed factors that have been shown to play a role in autoimmune diseases that share features with uveitis. One of the polymorphic genes that were recently shown to be involved in autoimmune disease such as inflammatory bowel disease, rheumatoid arthritis and diabetes belong to the so called family of Tumor necrosis factor (TNF)-receptor-associated factors (TRAF).TRAF5 is one of the factors belonging to the TRAF family and is significantly expressed in lung, thymus, spleen and kidney as well as in peripheral blood cells TRAF3IP2 is another member of the TRAF family and encodes Act1 protein, which binds to TRAF5 thereby playing an important role in controlling inflammatory responses and autoimmunity through its effect on CD40L and IL-17 signaling Based on earlier reports in RA, psoriasis and psoriatic arthritis All subjects gave their written informed consent for this study, and the study protocol was approved by the Ethics Committee of the First Affiliated Hospital of Chongqing Medical University, Chongqing, China (Permit Number: 2009-201008). All procedures followed the tenets of the Declaration of Helsinki.A case-control genetic study of two genes in individuals with or without uveitis was performed. 789 BD patients, 940 VKH patients and 1601 healthy unrelated individuals were included in this study. All subjects were of Han Chinese descent. Blood samples were obtained from the First Affiliated Hospital of Chongqing Medical University  and the Uveitis Study Center of the Sun Yat-sen University  through 2005 until 2013.The diagnosis of BD was based on the criteria of the International Study Genomic DNA samples of BD, VKH patients and healthy controls were extracted by using the QIAamp DNA Blood Mini Kit . Rs6540679, rs33980500, rs13210247, rs13190932 were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), the primers used to amplify the target DNA sequence by PCR are shown in 2 for 72 hours at a density of 1×106 cells/ml. Total RNA was extracted from the cultured PBMCs using TRIzol (Invitrogen), followed by reverse transcription using a transcriptase kit . Real-time quantitative PCR was performed to compare the mRNA expression of TRAF3IP2 and TRAF5 gene, using the Applied Biosystem 7500 Real-time PCR System and was determined using the SYBR Green I Assay kit (Applied Biosysterms) and normalized to β-actin mRNA. Relative expression levels were calculated using the 2−ΔΔCt method. The sense and antisense primers used in this experiment are depicted in Peripheral blood mononuclear cells (PBMCs) were prepared from heparinized blood by Ficoll-Hypaque density-gradient centrifugation. Isolated PBMCs were cultured in 24-well plates containing RPMI-1640 supplemented with 10% calf serum , 2 mM L-glutamine and 100 U/ml penicillin/streptomycin . Cells were left unstimulated or were stimulated with a cocktail of anti-CD3  and anti-CD28 antibodies  to mimic antigen presentation. Alternatively the cells were stimulated with LPS , at 37°C in humidified 5% COThe concentration of IL-8, IL-6 and TNF-α in cell culture supernatants was measured with the human Duoset enzyme-linked immunosorbent assay (ELISA) Development kit  according to the manufacturers' instructions.c) with the Bonferroni correction by multiplying with the number of analyses performed. For c equaled to a p value multiplied by 9  or 3 . A p c<0.05 was considered significant.Hardy-Weinberg equilibrium was tested using the χ2 test for goodness of fit and a p-value<0.05 was considered as a significant disequilibrium. The patterns of linkage disequilibrium (LD) of the tested SNPs were compared using Haploview . Allele and genotype frequencies were compared between patients and controls by the χ2 test or two-sided Fisher' exact test using SPSS . The p values were corrected , skin lesions (78.0%), genital ulcer (57.9%), arthritis (39.1%), positive pathergy test (24.5%) and hypopyon (22.3%). The VKH patients could be subdivided according to seven clinical features, including nuchal rigidity (17.9%), headache (50.4%), scalp allergy (10.4%), tinnitus (40.5%), alopecia (39.8%), poliosis (36.6%) and vitiligo (21.5%).c = 1.08×10−3, OR = 0.617; pc = 1.30×10−3, OR = 0.674, respectively), whereas a higher frequency of the GG genotype was observed  compared with controls. As to rs10863888/TRAF5, an increased frequency of the AG genotype was observed in BD patients , while a decreased GG frequency  was found. The frequency of the AG genotype and A allele of rs6540679/TRAF5 were remarkably increased , while the frequency of the homozygous GG genotype was markedly decreased  in BD patients.Significant differences between BD patients and controls were observed for four SNPs. The frequencies of the rs12569232/TRAF5 CG genotype and C allele were significantly lower in BD patients  while the frequency of the AG genotype and G allele was higher compared with controls   .c = 1.33×10−9, OR = 0.446;. pc = 9.45×10−12, OR = 0.442; pc = 5.65×10−11, OR = 2.339), whereas the AA genotype and A allele of rs6540679/TRAF5 were both decreased in VKH patients .Although obvious differences were found for the rs10863888 TRAF5 genotypes in VKH patients compared with healthy controls, they became non-significant after Bonferroni correction. The frequency of the CG genotype and C allele of rs12569232/TRAF5 was markedly lower in VKH patients as compared to controls. The frequency of the GG genotype was much higher  while the AG genotype was increased in VKH patients compared with normal controls   .Since the most significant association between TRAF gene polymorphisms and uveitis was found for TRAF5, we studied the effect of the different genotypes on the expression of TRAF5 under normal or inflammatory conditions. Real-Time PCR was performed to detect mRNA expression from PBMCs obtained from healthy controls. We genotyped 79 controls for the three SNPs of TRAF5 and then used Real-Time PCR to detect their TRAF5 expression at the mRNA level without stimulation. No difference could be detected under this condition . FollowiThe aforementioned result showed that different genotypes of rs6540679/TRAF5 could affect TRAF5 expression and therefore a further study was designed to investigate if different genotypes of rs6540679/TRAF5 could also affect the cytokine response of PBMCs following LPS stimulation. We measured the level of TNF-α, IL-6 and IL-8 (n = 22), which are important TRAF5 downstream factors In this study, we show that TRAF5 and TRAF3IP2 gene polymorphisms are associated with VKH syndrome and ocular BD in a Han Chinese population. Highest p values were observed for the association between uveitis and TRAF5, and we therefore focused on this SNP to investigate a possible functional association. This approach revealed that carriers of the GG genotype in SNP rs6540679/TRAF5 had a higher TRAF5 mRNA expression level and enhanced TNF-α and IL-6 secretion, compared to AA and AG carriers. Expression was not altered for the other TRAF5 SNPs. We did not perform a haplotype analysis since the sample size was too small.Our finding that the G allele of TRAF5 rs6540679 had a protective effect (OR = 0.629) in BD and a risk effect (OR = 1.235) in VKH suggests that the role of these factors depends on the context whereby they are involved during the process of inflammation. Similarly TRAF rs12569232 is most significant in VKH while rs6540679 is the most significant in BD. The functional studies performed until now have only addressed the mRNA expression and as yet no studies have addressed the effect of the gene polymorphisms on the binding characteristics of the factors and their final effect on for instance NF-kappa-B activation. Both uveitis entities we investigated in this study differ markedly as to the causative mechanisms and the role of TRAF family members for each disease might be different. BD is considered to be caused by an aberrant inflammatory response towards certain environmental triggers whereas VKH is an autoimmune disease directed against melanocyte antigens Taken together, the results confirm earlier studies showing a role for TRAF5 in the control of inflammatory responses and the increased risk of certain polymorphisms of this gene for autoimmune or autoinflammatory diseases  [17], [1VKH syndrome and BD are two of the most common uveitis entities in China TRAF5 and Act1, encoded by the TRAF3IP2 gene, have been implicated in the control of immune and inflammatory responses To ensure the validity of our data we took the following measures. First, the sample sizes of VKH patients, BD patients and normal controls were large enough to ensure an association analysis, and our sample was larger than previous reports Although our results suggested that TRAF5 and TRAF3IP2 are associated with the development of BD and VKH syndrome, it is still unknown how these SNPs exert their roles in these two diseases. The complex and redundant function of the members of the TRAF family and the context they are involved in may dictate their possible divergent roles in immune mediated diseases.Taken together, our study, for the first time, provides evidence for a role of TRAF5 and TRAF3IP2 polymorphisms in the development of BD and VKH syndrome.Table S1Clinical features of the investigated BD patients.(DOC)Click here for additional data file.Table S2Clinical features of the investigated VKH patients.(DOC)Click here for additional data file.Table S3Primers and restriction enzymes used for RFLP analysis of the TRAF5 and TRAF3IP2 gene polymorphisms.(DOC)Click here for additional data file.Table S4RT-PCR primers for genes.(DOC)Click here for additional data file.Figure S1TRAF5 mRNA expression levels in PBMCs without stimulation, according to the genotypes of the three SNPs . Data are presented as means ± SD.(TIF)Click here for additional data file.