Background: The association between paraoxonase 2 (PON2) gene polymorphisms and type 2 diabetes mellitus (T2DM) has been extensively investigated in the Chinese population with conflicting results. In this study, we systematically evaluated the association between PON2 Ser311Cys and Ala148Gly polymorphisms and T2DM risk by pooling all relevant studies.Methods: We searched PubMed, Embase, CNKI, and Wanfang databases for the studies. The strength of association was determined by the allelic, homozygous, heterozygous, recessive, and dominant genetic models and measured as odds ratio (OR) and 95% confidence interval (CI), under fixed- or random-effect models.Results: There was no significant association between PON2 Ser311Cys polymorphism and T2DM under any of the genetic models: allelic , heterozygous , dominant , recessive , homozygous . Similarly, no significant association was found in PON2 Arg148Gly polymorphism under any of the models: allelic , heterozygous , dominant , recessive , homozygous .Conclusions: The PON2 Ser311Cys and Ala148Gly polymorphisms were not associated with the risk of developing T2DM in the Chinese population. Diabetes is a major cause of mortality and morbidity worldwide, and it has become an important public health problem in China , 2. NatiPON2 gene contains eight introns and nine exons. It is polymorphic and several common single nucleotide polymorphisms (SNP) have been identified thus far. Currently, genetic variations in PON2 gene may be associated with a number of disorders, such as cardiovascular disease and T2DM (The paraoxonase 2 (PON2) gene encodes a member of the PON multigene family, which includes two other known members sharing approximately 65% sequence similarity at the amino acid level, PON1 and PON3, located adjacent to each other on the chromosome 7q21.3–22.1 in humans . PON1 anand T2DM –14.PON2 gene and the risk in developing T2DM has increased the focus on the Chinese population. In Qu's study, PON2 Ser311Cys gene polymorphism was found to be significantly associated with an increased risk of T2DM in a northern Chinese population  and Ala148Gly (rs11545942/rs12026) polymorphisms in pulation . In contl groups . In addil groups  failed tn et al.  also obsPON2 Ser311Cys and Ala148Gly gene polymorphisms and the risk of developing T2DM in the Chinese population.It is crucial to address this inconsistency among the currently published studies. In the present study, a comprehensive meta-analysis was conducted by pooling all qualified individual data from case-control studies to make a precise conclusion on the association between This study was performed in accordance with the guidelines of the Meta-analysis of Observational Studies in Epidemiology (MOOSE) statement .PON2 Ser311Cys and Ala148Gly gene polymorphisms and the risk of developing T2DM in the Chinese population. The search was performed in the databases in English and Chinese with proper keywords: paraoxonase 2 gene  or variations  in combination with T2DM . In addition, the references from relevant reviews or primary studies were hand searched to identify additional studies.The following electronic databases PubMed, Embase, Wanfang Data, and China National Knowledge Infrastructure (CNKI) were searched for all case-control studies published up to August 2017 on the association of PON2 gene and the risk of developing T2DM. (b) case-control studies, regardless of sample size. (c) studies providing the numbers of PON2 genotypes or alleles in case and control subjects. (d) the distribution of genotypes in the control groups met the Hardy-Weinberg equilibrium . Disagreement between two investigators regarding the eligibility of any study was resolved by the consensus of a third reviewer.Two investigators independently reviewed all studies for eligibility. The inclusion criteria were as follows: (a) study investigating the associations between Ser311Cys (rs6954345/rs7493) and Ala148Gly (rs11545942/rs12026) in Data extraction was independently performed by two reviewers. A consensus on all items of data extraction was reached by both reviewers. The following items were extracted from each study: the first author's name, publication year, region of the study, mean age and gender distribution of the participants, number of genotypes, distribution of alleles, sample size in case, and control groups.PON2 Ser311Cys gene polymorphism; G vs. A of PON2 Ala148Gly gene polymorphism), homozygous , dominant , heterozygous  and recessive  were performed.In our meta-analysis, 5 genetic models as the allelic  (The odds ratio (OR) and their corresponding 95% confidence interval (CI) were employed to assess the association of to 100%) . If therto 100%) , 23. ThePON2 Ser311Cys and Arg148Gly polymorphisms with T2DM, 89 articles were excluded after screening the abstract and title, and 44 articles were excluded after screening the full-texts. Finally, 15 articles were included. One article investigated the two polymorphisms in the same population. As a result, a total of 12 eligible studies were included for meta-analysis of PON2 Ser311Cys polymorphism , heterozygous , dominant , recessive , homozygous  , heterozygous , dominant , recessive , homozygous  . By contrast, as is shown in Table PON2 Arg148Gly polymorphism under all five genetic models .The results of the heterogeneity analysis in PON2 Ser311Cys polymorphism. The analysis showed that the studies conducted by Sun et al. , homozygous , recessive , heterozygous  and dominant  .Moreover, no evidence of publication bias was also detected by Egger's test  information available for Chinese population in the 1000 genomics database (https://www.ncbi.nlm.nih.gov/variation/tools/1000genomes/), we found that the minor allele frequency of PON2 Ser311Cys and Ala148Gly were the same each other: 0.20 in Southern Han Chinese and 0.18 in Beijing Han Chinese. In addition, Dasgupta et al. reported that PON2 Ser311Cys and Ala148Gly polymorphisms were in strong linkage disequilibrium (LD) with each other (r2 = 0.81)  . TherefoPON2 Ser311Cys polymorphism, there was substantial between-study heterogeneity under all five genetic models. Therefore, the Galbraith plot was employed to identify the outliers that could be the possible sources of between-study heterogeneity. The Galbraith plot analyses indicated that the studies conducted by Sun et al. (I2 values immediately decreased to 14% in an allelic genetic model and 0% in other four genetic models, and the P-values of the Q-test in all genetic models were >0.1. Moreover, the pooled ORs remained statistically insignificant in all genetic models, which demonstrated that our meta-analysis results were stable and reliable.In the meta-analysis of n et al. , Xu and n et al. , and Qu n et al.  were thePON2 Ser311Cys and Ala148Gly gene polymorphisms with diabetic nephropathy and retinopathy in Caucasian populations (PON2 genetic polymorphisms were not associated with diabetic nephropathy and retinopathy in Caucasians. In addition, numerous meta-analyses have been conducted to determine the association of PON2 Ser311Cys and Ala148Gly gene polymorphisms with the risk of developing other diseases, such as coronary heart disease (PON2 Ser311Cys and Ala148Gly gene polymorphisms and the risk of developing T2DM.One previous meta-analysis assessed the association of ulations . However disease –39, isch disease , 41 and  disease . This cuPON2 genetic polymorphisms in the susceptibility to T2DM. Other susceptibility genes such as PON1 and PON3 gene may interact with PON2 gene by gene-to-gene effect for T2DM (PON2 Ala148Gly polymorphism and obesity were found in the risk of T2DM (Our study has some limitations. First, some of the included studies were based on small sample size, which may have resulted in a decreased power to detect a significant difference in the distribution of genotypes or alleles between cases and controls. Second, the pooled OR is based on the crude OR in the original studies. Because we could not obtain enough raw data from individual studies, the pooled data were not adjusted by potential confounding factors such as gender, age, smoking status, body mass index and waist-hip ratio. Third, T2DM is a polygenic hereditary disorder. The current study only focused on the role of for T2DM , 43. Fou of T2DM . FinallyPON2 Ser311Cys and Ala148Gly gene polymorphisms did not have a significant association with the risk of developing T2DM in the Chinese population. A well-designed study, with consideration of gene-to-gene and gene-to-environment interactions, should be conducted in the future.In conclusion, our meta-analysis confirmed that HR and J-QL conceived and designed the study. J-QL, HR, S-LT, and M-ZL performed the search. HR, S-LT, and J-QL analyzed the data. J-QL, HR, and M-ZL contributed reagents, material, analysis tools. HR, S-LT, J-QL, and HB wrote and review the manuscript. HR and S-LT revised the manuscript. HR and J-QL reference collection, data management, statistical analyses, paper writing, and study design.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Elevated levels of serum homocysteine (Hcy) have been associated with cardiovascular diseases and endothelial dysfunction, conditions closely associated with erectile dysfunction (ED). This meta-analysis was aimed to assess serum Hcy levels in subjects with ED compared to controls in order to clarify the role of Hcy in the pathogenesis of ED.  Medline, Embase, and the Cochrane Library were searched for publications investigating the possible association between ED and Hcy. Results were restricted by language, but no time restriction was applied. Standardized mean difference (SMD) was obtained by random effect models. p < 0.0001. Subgroup analysis based on prevalence of diabetes showed significantly higher SMD in subjects without diabetes (1.34 (95% CI 1.08–1.60)) compared to subjects with diabetes (0.68 (95% CI 0.39–0.97), p < 0.0025 versus subgroup w/o diabetes).  A total of 9 studies were included in the analysis with a total of 1320 subjects . Pooled estimate was in favor of increased Hcy in subjects with ED with a SMD of 1.00, 95% CI 0.65–1.35, CRD42018087558. Results from our meta-analysis suggest that increased levels of serum Hcy are more often observed in subjects with ED; however, increase in Hcy is less evident in diabetic compared to nondiabetic subjects. This study is registered with Prospero registration number  Prevalence of erectile disorders increases steadily with aging, from 1 to 10% in men younger than 40 years up to 70–100% in men older than 70 years . It is wHowever, given the importance of hemodynamics in allowing adequate erection, it should come as no surprise that ED shares the same risk factors of many cardiovascular (CV) affections .μmol/l in serum Hcy levels, whereas Clarke et al.  OR (“erectile”[All Fields] AND “dysfunction”[All Fields]) OR “erectile dysfunction”[All Fields]) OR ((“penile erection”[MeSH Terms] OR (“penile”[All Fields] AND “erection”[All Fields]) OR “penile erection”[All Fields] OR “erectile”[All Fields]) AND (“physiology”[Subheading] OR “physiology”[All Fields] OR “function”[All Fields] OR “physiology”[MeSH Terms] OR “function”[All Fields])) OR  OR “sexual behavior”[All Fields] OR “sexual”[All Fields]) AND (“physiopathology”[Subheading] OR “physiopathology”[All Fields] OR “dysfunction”[All Fields])) AND (“homocysteine”[MeSH Terms] OR “homocysteine”[All Fields]). The search, which accrued data up to February 28th 2018, was restricted to articles written in English, Italian, or French and studies including human participants. The identification of relevant studies was performed independently by two of the authors  into two separate standard Excel templates which were then cross-checked by the other author. Extracted data included data sources, eligibility, methods, participant characteristics, and results. Heterogeneity in serum Hcy levels was assessed using I2 statistics. Even when low heterogeneity was detected, a random effect model was applied, because the validity of tests of heterogeneity can be limited with a small number of component studies. We used funnel plots and the Begg adjusted rank correlation test to estimate possible publication or disclosure bias ; however, undetected bias may still be present because these tests have low statistical power when the number of trials is small. Statistical analysis was performed independently by two authors (SA and CA) with the meta package on the statistical software R (version 3.4.2) [Data from retrieved study were extracted and entered twice by two authors (n 3.4.2) .Out of 30 retrieved articles, 9 were included in the study . The stuI2 in studies assessing serum Hcy in men with or without ED was 85.3% ). Serum Hcy was significantly higher among subjects with ED compared to controls )  suggested no publication bias (The 5–1.35)) . A funneion bias .p < 0.0025 versus subgroup w/o diabetes). Despite remaining higher in the subgroup with “mixed” prevalence of diabetes, heterogeneity is remarkably reduced in both other subgroups.Sensitivity analysis was then performed based on prevalence of diabetes. Five studies –28 had dThe results of our meta-analysis suggest that Hcy is significantly associated with ED, showing a mean increase of 1.00 standard deviation between controls and ED patients. Among patients with ED, sensitivity analysis has shown that when stratified for presence of diabetes, subjects without diabetes are more likely to have increased levels of serum Hcy compared to diabetic subjects; conversely, this association appears evident in patients with diabetes even for small variation of Hcy, or rather, it could be argued that in diabetic patients, a small increase of Hcy is enough to negatively affect erectile function. In fact, the difference between diabetic and nondiabetic patients might suggest that in the presence of comorbidities, small changes in serum Hcy might negatively be involved in erectile function, based on the premise that multiple risk factors might act in a synergistic, rather than additive, fashion , 34. ThiAs previously stated, HHcy is closely associated with endothelial dysfunction. Studies in animal models have proven that HHcy markedly inhibits NO formation in rabbit isolated corpus cavernosum . NO actsOur study has some limitations, most notably the small number of studies involved and the lack of a clear definition of ED. A single study  assessedResults from our meta-analysis suggest that increased levels of serum Hcy are more often observed in subjects with ED: based on existing literature on this topic, a causative role for HHCy as an independent risk factor for ED can be postulated, although confirmation would require interventional studies aimed to decrease serum Hcy levels considering erectile function as primary outcome. Actually, only in rat model of HHcy has been observed an improvement in erectile function after being treated with a demethylation agent . We also


BOLL gene  and intracranial aneurysm (IA) susceptibility. We aimed to perform an updated meta-analysis for the potential IA-susceptibility locus in large-scale multi-ethnic populations. We conducted a systematic review of studies identified by an electronic search from January 1990 to March 2019. The overall estimates of the “G” allele of rs700651, indicating IA susceptibility, were calculated under the fixed- and random-effect models using the inverse-variance method. Subsequent in silico function and cis-expression quantitative trait loci (cis-eQTL) analyses were performed to evaluate biological functions and genotype-specific expressions in human tissues. We included 4513 IA patients and 13,506 controls from five studies with seven independent populations: three European-ancestry, three Japanese, and one Korean population. The overall result showed a genome-wide significance threshold between rs700651 and IA susceptibility after controlling for study heterogeneity . Subsequent cis-eQTL analysis showed significant genome-wide expressions in three human tissues, i.e., testis (p = 8.04 × 10−15 for ANKRD44), tibial nerves (p = 3.18 × 10−10 for SF3B1), and thyroid glands (p = 4.61 × 10−9 for SF3B1). The rs700651 common variant of the 2q33.1 region may be involved in genetic mechanisms that increase the risk of IA and may play crucial roles in regulatory functions.Previous genome-wide association studies did not show a consistent association between the  EDNRA , SOX17 (8q11.23), CDKN2B-AS1 (9p21), CNNM2 (10q24.3), FGD6 (12q22), STARD13 (13q13), and RRBP1 (20p12.1) [The term unruptured intracranial aneurysm (UIA) refers to a localized outpouching of the cerebral artery due to a defect in the muscular layer ,2. The iKN2B-AS1 p21, CNNM), CNNM2 q24.3, FG), CNNM2 q24.3, FGBOLL, PLCL1, SF3B1, and ANKRD44 within the 2q33.1 region, was underpowered in multi-ethnic populations [GBA, ARHGAP32, OLFML2A, and SCARF1, the authors also reported two variants in Korean adults, rs6841581, near the 5′-untranslated region (UTR) of EDNRA (p = 6.5 × 10−4), and rs700651, in the intron of BOLL (p = 0.008). Although ethnic differences in genetic susceptibility are known to exist [In previous GWASs, the association of rs700651, mapped to several genes with IA, such as ulations . Specifiulations . While aulations  reportedto exist , sample to exist . Thus, wBOLL”, “boule homolog, RNA binding protein”, “PLCL1”, “phospholipase C like 1 (inactive)”, “rs700651”, “genetic”, and “GWAS”. The inclusion criteria for the studies were as follows: 1) saccular aneurysm studies, 2) studies for candidate genes or GWAS, and 3) IA studies in human subjects. The exclusion criteria were as follows: 1) animal studies; 2) dissection or fusiform aneurysms; 3) infection or traumatic aneurysms; 4) concomitant aneurysms with other cerebrovascular diseases, such as moyamoya disease or arteriovenous malformations; 5) studies without clear information on genotypes, OR, and 95% CI; 6) reviews, meta-analyses, or case reports. PubMed, Embase, and the HuGE Literature Finder electronic databases were searched for studies from January 1990 to March 2019 using the following keywords: “aneurysm(s)”, “intracranial aneurysm(s)”, “subarachnoid hemorrhage”, “genetic association”, “p-values, and covariates adjusted in the multivariate logistic regression model, were collected from seven association studies. Additional subgroup meta-analyses were performed according to ethnicity. The analyses were conducted using the Genome-Wide Association Meta-Analysis (GWAMA) software, (http://www.well.ox.ac.uk/gwama/) [2 statistics in order to detect potential publication biases across the individual studies. Forest plots were drawn to describe the previous results for rs700651 obtained from each study, as well as the overall effect sizes under the additive-effect model. Funnel plots were drawn to visualize potential publication biases. Those plots were constructed using the STATA software v.11.2 .The information on rs700651 analyzed under an additive genetic model, such as allele frequency, ORs, 95% CIs, /gwama/) . The ove2) was estimated for variants within the rs700651 variant position  ± 500,000 bp to detect transcription factor binding sites (TFBSs), splicing sites, and coding variants strongly correlated with this variant. Furthermore, the regional plot was used to identify 1) variants with putative regulatory potential via the RegulomeDB scoring system [https://ldlink.nci.nih.gov/) [We further investigated rs700651 and its neighbor variants using in silico functional analysis with fine mapping for the 2q33.1 region. Pairwise linkage disequilibrium  mih.gov/) .https://gtexportal.org/home/) [p-value of 5 × 10−8 and a suggestive significance threshold p-value of 1 × 10−5 in the cis-eQTL analysis, which are commonly used in GWASs.In subsequent analyses, we evaluated the genotype-specific expression (GSE) of rs700651 in 48 human tissues via cis-expression quantitative trait loci (cis-eQTL) analysis through the Genotype-Tissue Expression  portal (g/home/) . Violin Among nine eligible studies, four studies were excluded from the final analysis because they lacked the outcomes of interest  or were p = 0.0025, p = 1.05 × 10−8), with minimal heterogeneity (p = 4.83 × 10−5) compared with a meta-analysis of East-Asians with controlled population heterogeneity  (p = 3.60 × 10−5). However, the meta-analysis of the overall East-Asian populations did not demonstrate a statistically significant association with IA   were described under the fixed- and random-effect inverse variance models shown in ogeneity . In the  0.3749) , which c 0.0513) C, with a 0.0513) D.2) and MAFs greater than 40% in the East-Asian populations   which showed LD with rs700651 (r2 = 0.2894).Out of a total of 1599 variants within the 1Mbp region, 228 variants showed a pairwise LD with rs700651 (0.8 < rulations . These lANKRD44 (p = 8.04 × 10−15 in testis tissue) and SF3B1 . In particular, 8 of 10 genome-wide and/or suggestive cis-eQTLs were downregulated in multiple tissues (−0.59 < slope < −0.12), with the exception of two suggestive cis-eQTL PLCL1 gene expressions . However, the BOLL gene, that directly contains this variant in the intron region, showed an insignificant cis-eQTL in human tissues . Out of the total 618 genotypic cis-eQTL results for rs700651, three cis-eQTLs reached a genome-wide significance threshold in two eQTL genes, p = 1.05 × 10−8). To the best of our knowledge, this study is the first updated meta-analysis on the association between the 2q33.1 region (rs700651) and IA in a large-scale multi-ethnic population containing 18,019 individuals. Overall, the “G” allele of rs700651 consistently increased the risk of IA development in both European-ancestry and East-Asian populations. We confirmed the effect of ethnic similarity on this variant, including shared MAFs and overall effect sizes, in this study. Specifically, the overall results reached a genome-wide significance threshold in our meta-analysis controlled for study heterogeneity  gene family, is involved in germ cell development. Accordingly, it functions in the reproductive system, in particular, in human spermatogenesis and infertility [BOLL promoter has been observed in patients with colorectal or lung cancers. Kang et al. [BOLL promoter was enhanced in colorectal cancer tissue compared to normal control tissues. In addition, xenograft mice produced by engraftment with the human BOLL gene showed significantly higher tumor volumes after subcutaneous implantation, indicating an oncogenic role for BOLL. However, the role of BOLL in the development of IA is not well understood. A previous study found that females experienced higher aneurysm growth and subsequent rupture after menopause than premenopausal women, suggesting a possible hormonal effect on IA pathogenesis [BOLL encoded by the 2q33.1 region should be evaluated further. The ertility ,24. Aberg et al.  reportedogenesis . Kubo etogenesis  reportedogenesis . Furtherogenesis . Yeap etogenesis  reportedogenesis . AccordiThe initial GWAS of IA conducted in 2008 suggested a significant association between the “G” allele of rs700651 and IA in European and Japanese populations [p = 1.05 × 10−8). The 2q33.1 locus, a region harboring potential regulatory elements, showed regulomeDB scores in in silico functional analysis related to genetic mechanisms of IA formation via fine mapping. In addition, genotypic cis-eQTLs for rs700651 mapped to the regulatory regions of PLCL1, SF3B1, and ANKRD44 showed strong expressions in multiple human tissues, such as testis, tibial nerves, and thyroid glands (p < 5 × 10−8). If more studies are included in further meta-analyses, the reliability of this locus as a biomarker for IA susceptibility in multi-ethnic populations, as well as in ethnic subgroups, may be confirmed.There are some limitations in this study. The MAF of the “G” allele of rs700651 showed an approximate twofold difference in both European-ancestry and East-Asian populations , confirmed in the 1000 Genome Phase 3 reference panel. Despite the fact that the allele frequency difference in the general populations and a potential ethnic difference in human genome structures were present, we demonstrated a disease-specific genetic similarity in the ethnic populations tested, which showed the same effect direction of this variant susceptibility to IA. Our ethnic subgroup meta-analysis showed a lack of discriminative power to determine IA susceptibility variation in ethnically stratified GWASs due to the small effect size and the limited sample size of some populations, such as those of European-ancestry, East-Asian, and other populations. Nevertheless, our overall results highlighted strong power and a high genome-wide significance threshold (BOLL) and the regulation of synaptic transmission (PLCL1), related to IA susceptibility. Nonetheless, mapping heritability to a point variation is challenging, because single genetic variations cannot account for all heritable IAs and because cerebral aneurysms are affected by multifactorial interactions and components. Thus, the development of multiple polygenic models composed of IA-predicting variants should be pursued in future studies. From this point of view, our findings may offer better genetic information for IA diagnoses and provide a foundation for the next step in understanding IA pathogenesis.This meta-analysis highlights how an increase in the statistical power of rs700651 mapped to the 2q33.1 region played an important role in identifying regulatory mechanisms, including the positive regulation of translational initiation (


ADRB3) gene rs4994 polymorphism could affect the individual risk of childhood and adolescent overweight/obesity remains controversial. This meta-analysis was performed to estimate the prevalence of this polymorphism in overweight/obesity, and test the potential association by summarizing existing evidence. Comprehensive literature search in PubMed, Web of Science, Cochrane Library, Wanfang, and CNKI databases was performed to identify eligible data sets. Finally, 16 studies involving 5,147 overweight/obese cases and 7,350 non-obese controls were included for further synthetic analyses. Odds ratio (OR) and its corresponding 95% confidence intervals (CIs) were statistically calculated. Totally, 69.9% of the included subjects came from East Asia. In the meta-analysis for overall population, statistically significant associations with increased risk of childhood and adolescent overweight/obesity were identified in allele model , heterozygote model , and dominant model . Further stratified analysis according to geographical regions revealed that the statistical significance could only be detected in the East Asia subgroup in allele model, homozygote model, heterozygote model, and dominant model. In summary, our meta-analysis indicated that the ADRB3 rs4994 polymorphism could significantly increase the risk of childhood and adolescent overweight/obesity, especially for the East Asia’s population.Whether the Adrenoceptor Beta 3 ( Emerging as a serious health problem worldwide, childhood and adolescent obesity, a complex and multifactorial metabolic disorder, could lead to not only improper physical and mental development , but alsADRB3 rs4994 polymorphism (Trp64Arg), a T to C switch leading to the replacement of tryptophan by arginine at position 64, has been related to lower resting metabolic rate  from each database’s inception to 30 May 2019, in Chinese and English, was conducted. Terms for the ADRB3 gene and rs4994 polymorphism , definitions for single nucleotide polymorphism , synonyms for obesity , and keywords for ‘childhood and adolescent’  were merged in the Boolean expression for database query.A comprehensive literature search for relevant studies published on pre-reviewed journals in five databases [PubMed  Outcomes: obese or overweight. For publications with the same case-control population, only the largest or most complete study was included. Studies with the overall sample size less than 50 were excluded to avoid obvious sampling bias distorting the results.The eligibility of individual studies was evaluated by four investigators  who independently used the predefined exclusion/inclusion criteria. Case–control studies providing original genotype data comparing the allele frequency difference between obese case and control samples were selected. All included studies had to meet the following criteria: (1) Language: studies published in Chinese or English; (2) Participants: children or adolescents; (3) Exposure of interest: genotypes of the http://www.ohri.ca/programs/clinical_epidemiology/oxford.asp, accessed on 1 June 2019) was used to comprehensively assess the quality of included studies in this meta-analysis. The summarized scores for all subscale point items were used to categorize study quality as either low (<4), medium (4–6), or high (>6). Deviations from Hardy–Weinberg equilibrium (HWE) in control populations were assessed with an online calculator (http://ihg.gsf.de/cgi-bin/hw/hwa1.pl). P-values less than 0.05 indicate statistical significance for HWE deviation.A predefined data collection form was used for this review. The extracted data covered information regarding sample characteristics and features of study design. Author’s name, publication year, country, region, polymorphism detection method, definition method for obesity/overweight, and genotype data for case and control were recorded. The Newcastle–Ottawa Scale (NOS) quantification system . If no significance was detected, a fixed effect model (the Mantel-Haenszel method) would be used [P-value <0.05, the null hypothesis that there is no genetic impact of the ADRB3 Trp64Arg polymorphism on the increased risk of childhood and adolescent overweight/obesity could be rejected. Subgroup analyses were performed according to the geographical region (East Asia versus others), HWE status (consistent versus inconsistent and unassessable), sample size (no less than 200 versus less than 200), and sex . Publication bias was not only visually measured based on the extent of asymmetry of Begg’s funnel plot, and also statistically evaluated by Egger’s regression test [P-value <0.05 defines the statistical significance). If significant publication bias was detected, the Duval and Tweedie’s rank-based ‘trim-and-fill’ method was applied to impute for potentially missing data sets and adjust the effect of publication bias [All statistical analyses were performed using STATA Statistical Software . Initially, significance for heterogeneity between studies was evaluated using  be used . Otherwi be used . Odds raion test  and Beggion test   recommendations . Four inOf the 239 individual publications initially identified, 16 unique studies ,29,30 in2132p = 0.08) and dominant model (p = 0.05), so random effects model was used. Fixed effect model was adopted for synthetic analyses in allele model, homozygote model, and recessive model. Pooled estimates of overall and subgroup synthetic analyses were summarized in ADRB3 rs4994 polymorphism and obese risk was detected in allele model , heterozygote model , and dominant model , but not in homozygote model  and recessive model . Statistical significance for allele model, heterozygote model, and dominant model could be detected in both male and female subgroups. When stratifying the data sets according to the geographical region, we identified a statistically significant association between this polymorphism and obese risk in the subgroup of East Asia in allele model, homozygote model, heterozygote model, and dominant model . However, Egger’s regression test revealed there is slightly significant publication bias in the dominant model (p = 0.02), but not in the other models . The summary analysis incorporating four additional hypothetical studies using the trim-and-fill method continued to reveal a statistically significant association between the ADRB3 rs4994 polymorphism and risk of childhood and adolescent overweight/obesity . Begg’s rank correlation test for publication bias showed no statistical significance in all five genetic models (allele model: The leave-one-out sensitivity analysis iteratively removed one data set at a time to determine whether the statistical significance of the pooled estimates was driven by any single study. No statistically significant changes could be observed , suggestADRB3 rs4994 polymorphism on the risk of childhood and adolescent overweight/obesity despite previous case–control studies [There is still uncertainty regarding the genetic impact of the  studies . Our sysMeta-analyses pooling scattered epidemiological studies are generally prone to between-study heterogeneity and bias, which could partly impact the difficulty in drawing conclusions based on pooled estimates . In the This meta-analysis has several merits. Our study endorsed the recommendations by the PRISMA guidelines . The broADRB3 rs4994 polymorphism on the risk of overweight/obesity among children and adolescents are reassuring. There is a statistically significant association between the rs4994 polymorphism and overweight/obesity risk. However, this meta-analysis shows that the C allele of this polymorphism in the ADRB3 gene is a risk factor for overweight/obesity only in children and adolescents from East Asia.In conclusion, the results of this meta-analysis evaluating the genetic impact of the


TNF-α-308G>A and TNF-α-238 G>A polymorphisms and severity of dengue infection. However, the results are inconclusive and most studies had small sample sizes. The objective of this study was to summarize the evidence of association between TNF-α-308 G>A and TNF-α-238 G>A and severity of dengue infection.Individual studies have assessed the association between TNF-α-308 G>A or TNF-α-238 G>A and the risk of severe dengue infections. To determine the information size required for this meta-analysis study, a trial sequential analysis (TSA) was done. Eight studies (640 cases and 1275 controls), which assessed the association of TNF-α-308 G>A or TNF-α-238 G>A and the risk of DHF were included. Overall, we found no significant association between TNF-α-308 G>A and the DHF risk in the allelic model , the recessive model , the dominant model  or the additive model . There was also no significant association between TNF-α-238 G>A and DHF risk under the allele contrast model , the recessive model , the dominant model , or the additive model . On subgroup analysis, neither the Asian population nor the non-Asian population showed significant association between TNF-α-308 G>A/TNF-α-238 G>A and the DHF risk under any genetic models. Leave-one-out meta-analysis showed stability of the results. TSA plots suggested that the sample size in this meta-analysis study was below the required information size.This study follows the preferred reporting items for systematic reviews and meta- analyses of genetic association studies, recommended by PLOS One. We calculated pooled odds ratio and its 95% confidence interval (CI) to estimate the association between TNF-α-308/ TNF-α-238 G>A and the risk of developing severe dengue infection. Large studies with evidence of Hardy-Weinberg equilibrium, assessing gene-gene interactions are recommended.The findings suggest an inclusive evidence of the association between  Aedes aegypti and Ae. albopictus mosquitoesThe clinical manifestations of DENV infection vary from mild  to the severe form of dengue haemorrhagic fever (DHF). It is estimated that 2.5 billion people worldwide are at risk for DENV infection. About 5% of those at risk will have severe forms such as DHF and dengue shock syndrome (DSS), which are potentially fatal [Dengue fever is endemic in the tropics and sub-tropics and it is the most common arboviral infection caused by dengue viruses , which are transmitted primarily by the bite of The exact pathogenesis of progression from mild to severe forms of dengue infections is not known. A number of studies have suggested that leakage of plasma, which differentiates DHF from DF is attributed to the direct and indirect effects of cytokines or chemical mediators on the vascular endothelial cells –5. In anTNF-α gene, of which the two most frequently investigated are at position -308 [TNF-α and the increased risk of colorectal cancer under the homozygote model [There is an array of polymorphisms regulating the ion -308  and at -te model , and forte model .TNF-α-308 G>A or TNF-α-238 G>A promoter polymorphism and the risk of dengue infections in different populations are available in the literature. However, the results are inconsistent. Most of these studies are based on small sample sizes, with low statistical power. A meta-analysis of available individual genetic association studies is valuable to ascertain whether the polymorphisms of TNF-α-308 G>A and TNF-α-238 G>A gene affect the risk of severe dengue infections. Published meta-analyses in this field are also available, but there are concerns related to the selection of studies included [TNF-α-308 G>A or TNF-α-238 G>A and severity of dengue infection.Individual studies assessing the association between the included , the SNPincluded , the accincluded  and geneincluded . Taken tThe current study conformed to the checklist for meta-analysis of genetic association studies specified for PLOS One approach  S1 Tabl.TNF-α-238 G>A”. The details of search in PubMed database is provided in The potentially relevant studies were searched in the health-related databases of PubMed, Ovid Medline, google scholar and web of science and with the combination of the keywords with Boolean operators: “dengue” OR “DHF” OR “dengue haemorrhagic fever” OR “DSS” OR “dengue shock syndrome” AND “tumour necrosis factor- alpha-308” OR “rs1800629” OR “TNF-α-308 G>A” OR “tumour necrosis factor-alpha” OR “The search was restricted to the publications in English until January 2018. We also searched manually the references of articles retrieved and relevant systematic reviews to find any additional studies.TNF-α-308 G>A (including rs 1800629), TNF-α-238 G/A or both were investigated; (2) they were case-control design (retrospective or nested case-control) with an outcome of severe dengue infections (DHF/DSS) measured as incidence or prevalence, and (3) there was sufficient information to extract the genotype frequency both in cases and controls.Human studies of any sample size that assessed severe dengue infection were included, if (1) Severe dengue infection in the current analysis encompassed DHF and/or DSS as defined in the primary studies. Studies, which did not meet the inclusion criteria were excluded. Studies based on family or sibling-pairs were not considered.Two investigators independently screened the titles and abstracts, and then selected the relevant full-text articles, according to the inclusion criteria. The two investigators independently extracted data, using a piloted data extraction form. Information collected from each study included: first author, publication year, country, study setting , the frequency of cases and controls, the racial descent (Asian or non-Asian), method of genotyping and genotype/allele frequencies in cases and controls. If allele frequency was zero, then we added 1 to all allele, following the Laplace approximation . Any disThe two investigators independently evaluated the methodological quality of studies with the use of the Newcastle-Ottawa Scale (NOS) . These sp>0.05) [An evidence of HWE in the control population was assessed (if not provided in the study included) or re-assessed  in this meta-analysis with the use of the goodness-of-fit test (p>0.05) .TNF-α-308 G>A or TNF-α-238 G>A and the risk of developing DHF/DSS was estimated using odds ratios (ORs) and its 95% confidence intervals (CIs) for individual studies included. Heterogeneity was evaluated with the use of I2 test. The I2 test values indicates the percentage of total variation across studies that is attributable to the heterogeneity rather than chance. I2 values lie between 0% and 100%, with greater than 50% regarded as having substantial heterogeneity [Following a method described , the strogeneity . For pooogeneity . All eigogeneity ,22.Trial sequential analysis (TSA), a tool for adjustment of random error risk, was done for estimation of the required information size . It is cmeta package in R 3.4.3 software (The R Foundation), respectively. TSA plot was done with TSA software .Forest plots and leave-one-out meta-analysis were done with RevMan 5.3  and TNF-α-308 G>A. Only 3 studies assessed both TNF-α-308 G>A and TNF-α 238 G>A [ 238 G>A ,30,32 in 238 G>A –43 were TNF-α-308 G>A and TNF-α-238 G>A gene polymorphisms. Two studies each were from Brazil [m Brazil , 31 and m Brazil ,32, whilm Brazil , India [m Brazil , Malaysim Brazil . Study sm Brazil  to 196 [m Brazil , 26. AllTNF-α-308 G>A and the DHF risk under the allelic model , the recessive model , the dominant model  and the additive model . On stratification, the Asian population or the non-Asian population showed no significant relationships between TNF-α-308 G>A and the DHF risk under any genetic models, except a recessive model with the Asian population   .TNF-α-238 G>A, overall there was no significant association with the DHF risk in the allelic model , the recessive model , the dominant model  and the additive model . Also, a stratified analysis with the Asian population or the non-Asian population did not show significant association between TNF-α-238 G>A polymorphism and the risk of DHF in any genetic models   . This wap <0.05, even the required information size of 4059 is obtained , suggesting that TNF-α-308 G>A was not associated with DHF [A published meta-analysis has documented that otection . The curwith DHF . AlthougThe lack of association either protective or enhancement of the DHF risk in the current analysis might be due to insufficient number of participants, which is supported by the TSA plot. It showed that the included eight studies in this meta-analysis could only contribute to 65.7% of the required information size. This implied there was insufficient information to provide conclusive results.TNF-α in the development of DHF risk. In animal models, IFN-γ does not directly induce plasma leakage, but promotes TNF-α production through activated monocytes [TNF-α-308 G>A and other cytokines are important to establish the significant role in the DHF risk. This was supported by a study in Malaysia that showed a significant correlation between interactions of TNF-α-308G>A genotype and IL10 non-GCC haplotypes, IL12B pro homozygotes  and IL-12B 3’UTR AC and the protective effects against DHF/DSS [TNF-α, IFN- γ and activated complement proteins enhance plasma leakage of endothelial cells in secondary dengue infection [TNF-α-308 G>A and the risk of severe dengue. This implied that the criteria for dengue severity had no impact on the direction of association.Another possible reason of a lack of association in the present analysis may be due to the limited role of onocytes  and inteonocytes . As suchnfection . A smallTNF-α-238 G>A or TNF-α-308 G>A and other genes (gen-gen interaction/synergism) or other potential confounding factors such as nutritional status of the patients included [We acknowledge the limitations of the present review. Approximately 75% of the global population residing in the Asia Pacific Region are exposed to DENV . Only 36included . Hence, There are some strengths in this meta-analysis compared with other reviews in this field. To be comprehensive, we have added more than one SNP in the present analysis. We performed this review, following the guideline for meta-analysis of genetic association studies . MoreoveS1 Table(DOC)Click here for additional data file.S2 Table(DOC)Click here for additional data file.S3 Table(DOC)Click here for additional data file.S4 Table(DOC)Click here for additional data file.S5 Table(DOC)Click here for additional data file.


Hepatocellular carcinoma (HCC) is the sixth-most common malignancy worldwide. Multiple previous studies have assessed the relationship between TM6SF2 gene polymorphism and the risk of developing HCC, with discrepant conclusions reached. To assess the association of TM6SF2 rs58542926 T/C gene polymorphism with liver cancer, we performed the current meta-analysis.This study queried the MEDLINE, PubMed, EMBASE, and CENTRAL databases from inception to April 2019. Case-control studies assessing the relationship between TM6SF2 rs5854292 locus polymorphism and liver cancer were selected according to inclusion and exclusion criteria. The Stata 12.0 software was employed for data analysis.A total of 5 articles, encompassing 6873 patients, met inclusion criteria and were included in the meta-analysis. Statistical analysis showed that the TM6SF2 gene polymorphism was significantly associated with liver cancer in the allele contrast, dominant, recessive and over dominant models . The Egger’s test revealed no significant publication bias.The present findings suggest a significant association of TM6SF2 gene polymorphism with HCC risk in the entire population studied. Hepatocellular carcinoma ranks as the sixth-most common malignancy worldwide . Recent The current meta-analysis complied with Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines . The seaA comprehensive search for literature addressing the genetic associations of TM6SF2 variants in patients with HCC was conducted in the Medline, EMBASE, PubMed and CENTRAL databases without language restriction, from inception to April 2019. The specific search strategy was “ Liver Neoplasms or Hepatic Neoplasms or Hepatocellular Cancer or Liver Cancer” and “TM6SF2 protein or Transmembrane 6 superfamily member 2”. Table Relevant articles were initially selected based on title and abstract. Then, two authors reviewed the full texts to select qualified articles based on set eligibility criteria. Any disputes during the selection process were discussed with and resolved by a third investigator.Inclusion criteria:(1) the study cohorts included TM6SF2 rs58542926 T/C gene polymorphism in patients with liver cancer and non-hepatoma individuals; (2) histological features were assessed by liver biopsy, and diagnostic criteria were clearly stated; (3) case-control studies were enrolled, and the control group included non-hepatoma cases; (4) if two (or more) studies included the same cohort, the most recent was included to avoid repeated statistics; (5) the risk ratios of hazardous variants on the susceptibility of hepatocellular carcinoma were reported or could be calculated; (6) the full text could be retrieved by different ways. Exclusion criteria: (1) the source of enrolled cases in the article is unclear;(2) no clear diagnostic criteria for HCC described; (3) data collection and analysis methods unscientific or inappropriate; (4) lack of detailed genotyping data; (5) no-case-control study; (6) in animal studies.Two experienced authors independently extracted the necessary data and information from eligible publications according to a predetermined data extraction form. The information extracted from all the selected studies included: first author’s surname, publication year, country in which the study was conducted, total numbers of patients in the case and control groups, sex ratio, age, and body mass index (BMI), as well as the numbers of cases and controls with the C/C, C/T, and T/T genotypes. Whether genotype distribution was consistent with the Hardy-Weinberg equilibrium (HWE) was also recorded.P < 0.05 considered to present statistical significance.The Egger’s test was used for assessing publication bias, with χ2 test, and P > 0.05 was regarded as consistent with the HWE. The random or fixed effects model was used to pool ORs based on heterogeneity assumption, Heterogeneity across studies was determined by the Q- and I2 tests. The fixed-effects model was used in the case of nonsignificant heterogeneity ; otherwise, the random-effects model was utilized. In order to explore the effect of a single study on overall results, sensitivity analysis was performed by removing one study sequentially to evaluate its effect on the overall results under all genetic models. The STATA 12.0 software was employed for statistical analyses.The association of the T/C polymorphism in the TM6SF2 gene with HCC susceptibility was evaluated by calculating pooled odds ratios (ORs) alongside 95% confidence intervals (CIs) in the allelic, dominant, recessive, and super-dominant models. The HWE for each study was measured by the There were a total of 79 relevant studies compliant with the strategy, of which 24 were excluded as duplicates. After further title and abstract review, 33 reports were excluded as irrelevant to this meta-analysis. The second-round of review was based on careful full-text review of the 22 retained papers. Then, 17 reports were exclude, leaving 5 that were included in the final analysis. Figure Table Five studies included in the current meta-analysis described the association of TM6SF2 rs58542926 T/C gene polymorphism with susceptibility to liver cancer. The allelic (T vs C), dominant (CT + TT vs CC), recessive (TT vs CT + CC), and super-dominant (CC + TT vs TC) models were assessed. Since one of the studies provided no specific frequency distributions for CC, CT, and TT, and only provided frequency distribution in the dominant model (CC vs CT + TT), the five reports were included for assessment in the dominant model. In the allelic, recessive and super-dominant models, 4 studies were included, with a total of 2462 HCC patients and 3464 controls. The fixed effects model was employed for pooled ORs since nonsignificant heterogeneity was detected. The results showed that the TM6SF2 gene polymorphism was significantly associated with susceptibility to liver cancer , sensitivity analysis suggested that the result was relatively stable. There are two articles on the relationship between TM6SF2 gene polymorphism and HCC caused by viral hepatitis. The results of META analysis on the dominant gene model showed that the pooled risk of liver cancer was higher in the TT + CT genotype compared with the CC genotype , sensitivity analysis suggested that the result was relatively robust. There is one article to study the relationship between TM6SF2 gene polymorphism and NAFLD-induced HCC, so there is no META analysis result. Due to the limitation of the number of articles included, the results of stratified analysis of HCC by cause are not satisfactory, but according to the current analysis results, the cause of HCC seems to have no effect on the meta analysis. There are also some documents that confirm this claim. Stickel F et al. showed that the development of HCC was independently associated with TM6SF2 rs58542926, Carriage of TM6SF2 rs58542926 is an independent risk factor for the development of HCC in people with alcohol-related cirrhosis [To further clarify whether the different causes of liver cancer affect the results of the meta-analysis, we divided the causes of HCC into viruses, NAFLD, and alcoholic liver disease. There are 3 articles on the relationship between TM6SF2 gene polymorphism and HCC caused by alcoholic liver disease. The results of META analysis on dominant gene model showed that the pooled risk of liver cancer was higher in the TT + CT genotype compared with the CC genotype , dominant , recessive  and super-dominant  models  features a guanine to adenine substitution (nucleotide position 499), resulting in glutamate to lysine change at amino acid position 167 (E167K) . SubcellRecent studies have shown thatTM6SF2 may not solely be a marker associated with increased risk of HCC, but may be involved in the development of HCC at the cellular level. Shuixian Du et al. investigated the effect of TM6SF2 E167K on the expression levels of TNF-α, IL-2, IL-6 and IL-8 in the HCC cell HEPA 1–6,and demonstrated that overexpression of the TM6SF2 E167K protein significantly up-regulates the expression of IL-2 and IL-6 . Their fAlthough the relationship between TM6SF2 gene polymorphism and the risk of liver cancer has attracted attention from many researchers, results vary from study to study. In a single study, the stability and reliability of the research results are affected by the limited sample size. However, meta-analyses use suitable mathematical models to perform quantitative analysis of multiple identical or similar research results, increasing the test efficiency of research results.Here, a search strategy was designed, and article quality was assessed based on the Oxford Critical Appraisal Skill Program guidelines  . ArticleIn sensitivity analysis, the meta-analysis findings were relatively stable. Publication bias is an important factor affecting the results of a meta-analysis. The Egger regression method was used to demonstrate that the meta-analysis had no overt publication bias, suggesting that the above results were reliable.The limitations of this study could not be ignored. Firstly, due to the limited number of articles included, this meta-analysis failed to stage the tumors for group discussion. Secondly, this meta-analysis only involved single factor studies,the interactions of TM6SF2 gene polymorphisms and environmental factors, obesity, alcohol intake, intake of the fungal metabolite aflatoxin, and hepatitis B and C infections were not taken into consideration . MeanwhiIn summary, TM6SF2 rs58542926 gene polymorphism is significantly associated with liver cancer susceptibility. It should be further investigated whether the TM6SF2 rs58542926 variant could be screened for early diagnosis of liver cancer.Additional file 1: Figure S1. Sensitivity analysis of TM6SF2 rs58542926 T/C in the dominant model. Figure S2 Sensitivity analysis of TM6SF2 rs58542926 T/C in the allelic model. Figure S3. Sensitivity analysis of TM6SF2 rs58542926 T/C in the recessive model. Figure S4. Sensitivity analysis of TM6SF2 rs58542926 T/C in the super-dominant model. Figure S5. Egger’s funnel plot of TM6SF2 rs58542926 T/C in the dominant model. Figure S6. Egger’s funnel plot of TM6SF2 rs58542926 T/C in the allelic model. Figure S7. Egger’s funnel plot of TM6SF2 rs58542926 T/C in the recessive model. Figure S8. Egger’s funnel plot of TM6SF2 rs58542926 T/C in the super-dominant model.


Conflicting results have been reported regarding differing studies on the association between T-cell immunoglobulin and mucin domain 3 polymorphisms and autoimmune disease. The purpose of the present study was to evaluate the association of TIM-3 rs1036199 (4259 G/T) polymorphism with autoimmune disease susceptibility. A meta-analysis was performed to obtain a more precise evaluation of the association. Ten eligible studies were retrieved by searching PubMed, Embase and Web of Science databases, and statistical analyses were performed using STATA software. The pooled results indicated that TIM-3 rs1036199 polymorphism was significantly associated with an increased risk of overall autoimmune disease in allele comparison  and heterozygous comparison . Subgroup analyses based on disease type demonstrated that TIM-3 rs1036199 polymorphism was associated with an increased risk of rheumatic arthritis , especially in Asian populations. Autoimmune diseases (ADs) are a major health issue worldwide, affecting nearly 10% of the population . It has + T cells and at a low level on Th17 cells [+ T cells [TIM-3 is a negative regulator of immune responses and can be expressed on activated Th1 cells, CD817 cells . These c17 cells ,17. Bloc T cells . TIM-3 a T cells ,20.Several studies have demonstrated that TIM-3 polymorphism could be associated with ADs, such as multiple sclerosis (MS), Graves’ disease (GD), Hashimoto’s disease (HD), autoimmune thyroid diseases (AITDs), ankylosing spondylitis (AS), idiopathic thrombocytopenic purpura (ITP), systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) . ResultsFor the meta-analysis, PubMed, Web of Science and Embase databases were searched without language limitations. The final search was performed on July 7, 2018, using search terms: ‘T cell immunoglobulin and mucin domain 3 OR TIM-3 OR 4259 G/T OR rs1036199’, ‘autoimmune diseases OR autoimmunity’ and ‘polymorphism OR variant’. In order to identify additional eligible studies, references from all relevant articles were also included.Selection criteria were independently assessed by two researchers, based on several factors. First, assessment of the association between TIM-3 rs1036199 and ADs; second, only case–control studies were included, and finally that sufficient genotype data were available for odds ratio and confidence interval calculations. Agreement was reached via discussion in the case of any conflicts.Data were extracted from the eligible studies by two independent researchers. The following data types were extracted: the first author, publication year, country, ethnicity, disease type, genotyping method, source of control, total number of cases and controls, Hardy–Weinberg equilibrium and amount of cases and controls for every genotype. Any dispute was resolved by discussion.P<0.05 considered as a statistically significant disequilibrium. Odds ratio (OR) and 95% confidence interval (CI) were calculated to assess the association between TIM-3 rs1036199 polymorphism and ADs. Pooled OR results were derived from the combination of each study through comparison in allelic (G versus T) and heterozygote (GT versus TT) models. A Z-test was used to determine pooled OR results, and a P value <0.05 was considered significant. Heterogeneity was assessed using a standard Q-statistic test and an I2 test was used to quantify inconsistency. If the P-value of the Q-test was less than 0.1, or an I2 value >50%, ORs were pooled according to the random-effective model. Otherwise, the fixed-effective model was employed. Sensitivity analyses were conducted toward every genetic model to assess the influence of each individual study on combined ORs by sequentially deleting each study. Additionally, subgroup analysis was stratified by disease, ethnicity and source of control. Publication bias was evaluated by Egger’s test and Begg’s funnel plots. All statistical analyses were performed using STATA software, version 14.0 .The Chi-squared test was used to examine the Hardy–Weinberg equilibrium within the control group, with The study selection process is illustrated in P=0.142, I2 = 33.3%), but slight heterogeneity was found in allele model . Subsequently, subgroup analysis was conducted by ethnicity, source of control and disease type. When subgroup analysis was performed based on ethnicity, significant correlation was detected between rs1036199 polymorphism and increased risk of ADs in Asian populations .Begg’s funnel plots were performed to assess any possible publication bias, and no obvious asymmetry evidence was found according to the shape of the funnel plots A,B. SubsTIM-3 gene is located on chromosome 5q33.2 and is mainly expressed in Th1 cells . The TIMThe 4259 G/T polymorphism is located within exon 3 of TIM-3. A switch from T to G leads to the amino-acid substitution of arginine by leucine . The effTo the best of our knowledge, no previous meta-analysis has comprehensively investigated the association between rs1036199 polymorphism and AD risk. Our analysis revealed that TIM-3 rs1036199 polymorphism was significantly associated with an increased overall risk of AD. The AD risk was markedly more pronounced in Asian populations using allelic and heterozygous genetic models. Subgroup analyses based on disease type further revealed that TIM-3 rs1036199 polymorphism was only associated with an increased risk of rheumatoid arthritis and might have no effect on GD or ITP.There are, however, some limitations to the meta-analysis. First, for several ADs, the sample of studies is small, which may lead to insufficient power to detect a slight association. Second, most of the investigations analyzed were conducted on Asian populations, so further investigation into other ethnic populations is required. In addition, more eligible investigations on different kinds of autoimmune disease are recommended.In conclusion, our analysis indicates that TIM-3 rs1036199 polymorphism increases the susceptibility to AD in the overall population and in Asian populations. In particular, it shows that the TIM-3 rs1036199 polymorphism is associated with an increased genetic susceptibility to rheumatoid arthritis.


The inflammatory potential of diet has been shown to have an association with the risk of several cancer types, but the evidence is inconsistent regarding the related risk of urologic cancer (UC). Therefore, we conducted the present meta-analysis to investigate the association between the inflammatory potential of diet and UC.PubMed, Embase and Web of Science were searched up to July 31, 2018. Two reviewers independently selected the studies and extracted the data. The pooled risk ratio (RR) and its 95% confidence interval (CI) were calculated using the Stata12.0 software package.Nine case-control studies and three cohort studies including 83,197 subjects met the inclusion criteria. The overall meta-analysis results showed that individuals with the highest category of DII (dietary inflammatory index) were associated with an increased risk of prostate cancer ; subgroup analysis showed consistent results. For kidney and bladder cancer, significant positive associations were found in individuals with the highest category of DII score; however, no significant association was found between DII and the risk of urothelial cell carcinoma (UCC).Available data suggest that more pro-inflammatory diets are associated with an increased risk of prostate cancer, kidney cancer and bladder cancer. However, further well designed large-scaled cohort studies are warranted to provide more conclusive evidence. Prostate cancer, bladder cancer, and kidney cancer, the most common urologic tumors, are leading causes of cancer-related morbidity and mortality worldwide , 2. DespThe DII score, a literature-derived population-based dietary score, was developed to estimate the inflammatory potential of nutrients and foods in the context of a dietary pattern , 14. TheThe included studies were searched from PubMed, Embase and Web of Science up to July 31, 2018. Search terms were as follows: “ AND  AND (cancer OR carcinoma OR neoplasm).” In addition, a manual search of references in relevant articles was conducted to find other eligible studies. The search strategy flowchart is shown in Only studies meeting the following inclusion criteria were eligible: 1) studies with full text articles; 2) studies that reported the association between the inflammatory potential of diets and UC risk; 3) studies with odds ratios (ORs), relative risks (RRs), or hazard ratios (HRs) with corresponding 95% confidence intervals (CIs); and 4) the published language was English. Exclusion criteria were as follows: (1) studies referred to outcomes other than UC; (2) studies with partially unusable data; (3) review articles, meta-analyses, animal studies, conference abstracts or editorial articles.A quality assessment of the included studies was evaluated by the Newcastle-Ottawa Scale (NOS) . The NOSData extraction was independently conducted by two authors using a collection form that was checked by a third author. Disagreement was resolved through discussion and consensus finding. For each study, we collected the following information: (1) the first author’s name, year of publication, country, ethnicity, and sample size of the study; (2) mean age or age range, study design, and cancer type; (3) dietary assessment method, most fully adjusted risk estimate, and cofounders included in the final models.2 metric were used to assess heterogeneity among studies. The heterogeneity was considered significant when p<0.10 and I2>50%. Given that the included studies were conducted at a global level and addressed different types of cancer, random effects model was used to get more conservative results. Subgroup analyses based on study design and sample size were performed for prostate cancer. The significance of the summary OR was determined by the Z-test, and P<0.05 was considered as statistically significant. Begg’s, Egger’s test and funnel plots were used to assess potential publication bias. Sensitivity analysis was performed by excluding one study each time to evaluate the stability of the results. Statistical analysis was performed using STATA software version 12.0 .The multivariable-adjusted HR or OR with 95% CI for the highest versus the lowest DII score were pooled using random effects models. A Chi-square-based Q test and the IA total of 855 results were retrieved through literature searching. Of these, 843 studies were excluded based on inclusion and exclusion criteria. Finally, twelve studies considering 83,197 subjects met the inclusion criteria –30. Four2 = 42.2%). Additionally, publication bias was observed from the Begg (P = 0.013) and Egger regression tests (P = 0.019), as well as the funnel plot . The pooled RR for the highest versus lowest DII score was 1.46 (95% CI: 1.16–1.85)  .One study including 1,355 participants evaluated the association of DII score with bladder cancer risk. The pooled RR for the highest versus lowest DII score was 1.97 (95% CI: 1.28–3.03). One cohort study with 37,442 participants evaluated the association of DII score with urothelial cell carcinoma risk. The pooled RR for the highest versus lowest DII score was 1.24 (95% CI: 0.90–1.70) .Sensitivity analysis was performed for prostate cancer by omitting one study each time; the results showed that the overall pooled RRs were not influenced by any individual study , suggestThis is, to our knowledge, the first systematic review with a meta-analysis that evaluates the association between the inflammatory potential of diet and UC risk. Eleven studies with a total of 83,197 participants met the inclusion criteria and were finally included in the meta-analysis. The results showed that more pro-inflammatory diets, estimated by a higher DII score, are associated with an increased risk of prostate cancer, kidney cancer and bladder cancer.The etiology of urologic cancers  is complicated, and several risk factors are involved in their development and progression; in addition to environmental and genetic risk factors, lifestyle risk factors, such as dietary habits, also play important role in cancer development and progression –35. TherIn the present meta-analysis, a stronger association was detected between higher DII score and prostate cancer risks in the overall analysis. The pooled adjusted risk ratio (RR) for the highest DII score versus the lowest category was 1.62 (95% CI: 1.30–2.02). Subgroup analyses based on study design and sample size showed consistent results. Vázquez-Salas et al. reported that there is no evidence of an association between a pro-inflammatory diet and prostate cancer risk, in contrast to the conclusions of previous studies , 22, 25.When interpreting the results of the current study, some limitations should be considered. First, only two studies reported the relationship between the DII score and kidney cancer risk; additionally, one study for bladder cancer and one study for urothelial cell carcinoma reported a relationship. The sample size of included published articles was small, so sufficient data was unavailable. Second, the DII score was calculated by self-report, inevitably leading to some recall bias. Third, substantial heterogeneity reporting on prostate cancer was observed among studies; this may a result of the different number of food parameters, geographical region and follow-up duration. Finally, some publication bias exists in the results which may due to the limited studies in the present meta-analysis.This study suggests that a pro-inflammatory diet is associated with an increased risk of prostate cancer, kidney cancer and bladder cancer. Nevertheless, more large-scale, well-designed studies are needed to investigate the findings, and future research is needed to investigate whether an anti-inflammatory dietary pattern could constitute a beneficial nutritional choice for the primary prevention of UC.S1 Checklist(DOC)Click here for additional data file.S1 Fig(DOC)Click here for additional data file.


Circulatory osteocalcin (OC) has been widely used as a biomarker to indicate bone turnover status in postmenopausal osteoporosis (PMO). However, the change of serum OC (sOC) level in PMO cases compared to postmenopausal controls remains controversial.We searched the online database of PubMed and Cochrane Library. A meta-analysis of case-control studies was performed to compare the pooled sOC level between PMO patients and postmenopausal controls. Subgroup analysis according to potential confounding factors (different OC molecules and regions of the study population) was also performed.p = 0.28] between PMO patients and controls. Subgroup analysis also revealed no significant difference in intact OC  or N-terminal mid-fragment of the OC molecule  between groups. For different regions, no significant difference in sOC was found in Asian population between cases and controls , while the pooled sOC level was significantly higher in European PMO cases than controls .Ten case-control studies with 1577 postmenopausal women were included in this meta analysis. We found no significant difference in the pooled sOC level [mean difference (MD) = 1.84, 95% confidence interval (CI): , Our analysis revealed no significant difference in sOC level between PMO cases and controls according to all the current eligible studies. OC molecules are quite heterogeneous in the circulation and can be influenced by glucose metabolism. Therefore, sOC is currently not a good indicator for the high bone turnover status in PMO. More trials with standardized methodologies for the evaluation of circulatory OC are awaited to update our current findings. Osteoporosis (OP) is a systemic skeletal disorder characterized by low bone mass and deteriorated microarchitecture of bone tissue. The osteoporotic bone is fragile and susceptible to fracture . PostmenOsteocalcin (OC), also known as the bone Gla protein (BGP), is a 5.8 kDa, hydroxyapatite-binding protein that could be synthesized by osteoblasts, odontoblasts and hypertrophic chondrocytes  and is tAlthough serum osteocalcin (sOC) has been widely used as a bone turnover marker to indicate the high bone turnover status in PMO –9, the cTo obtain the evidence, we performed a meta-analysis of the available case-control studies to compare the sOC level between PMO patients and age-matched postmenopausal controls.We followed the Preferred Reporting Item for Systemic Review and Meta-analysis (PRISMA) , 15 and We searched PubMed and Cochrane Library online for all the available case-control studies published up to (12 October 2018), without restriction to regions and languages. Detailed strategy was:  AND . Conventional searches were supplemented by manual searches of the reference lists of all the relevant studies, review articles and conference abstracts.Studies were included if they met the following inclusion criteria: 1.case-control study design; 2.patients in the case group were diagnosed of PMO, according to the diagnostic criteria recommended by WHO ; 3.partiTwo literature reviewers evaluated the eligibility of potential titles and abstracts independently. Included studies were reassessed as full text by inclusion and exclusion criteria. Disagreement was solved by discussion. Further adjudication of a third reviewer was performed if the disagreement remained. The following data were then extracted from each eligible study: the first author’s name, year of publication, demographic information , sampling condition, manufacturer of the OC assay kit, target fragment of the sOC molecule and sOC level.Assessment to the risk of bias of case-control studies was performed using the Newcastle-Ottawa Scale (NOS) , as recop ≤ 0.10 (or I2 ≥ 50%), and homogeneous data was indicated by p > 0.10 (or I2 < 50%). A Fixed Effect model was used for homogeneous data, whereas a Random Effects model was used for heterogeneous data. Continuous variables of sOC level were reported with 95% confidence interval (95% CI) of the mean difference (MD) and p value. The test of overall effect with a p < 0.05 was considered to be statistically significant. Collected data were carefully inputted, and then rechecked by two reviewers respectively.The pooled sOC level was compared between cases and controls. Statistical analysis was performed using Review Manager (RevMan 5.3) software. First, heterogeneity of the included studies was tested. Heterogeneous data between studies was indicated by p ≤ 0.10 (or I2 ≥ 50%) and homogeneous data between subgroups was indicated by p > 0.10 (or I2 < 50%).Subgroup analysis according to different fragments of the sOC molecules and different regions of the study population was also performed, with the same method mentioned above. Heterogeneous data between subgroups was indicated by Six hundred and ninety five references were identified, of which 10 studies –27 fulfiCharacteristics of the included studies are summarized in Table Quality assessment of the included studies was summarized in Table 2 = 100%). We therefore pooled the sOC level from all the included studies with Random Effects model. A trend of increased sOC level in PMO cases could be observed, while the difference in sOC level between cases and controls was not statistically significant  , p = 0.84] between cases and controls , p = 0.98] , p = 0.28] to . This result indicated that the pooled test performance of this analysis could be influenced by a single study. Therefore, the conclusion should be carefully drawn and further tested.We removed one study at a time and re-compared the pooled sOC level between cases and controls. Significant change occurred when removing Al-Daghri’s study , which cSerum OC has been widely applied to indicate bone turnover status in PMO management with several advantages such as bone specificity, non-invasiveness and dynamic-response. However, updated knowledge of OC challenges its traditional role as a bone turnover indicator in vivo, and studies have reported conflicting results of the change of sOC level in PMO cases.PMO presents a high bone turnover rate with both increased bone resorption and formation –12. Accop = 0.28], which is not statistically significant. Subgroup analysis according to different regions and sOC fragments also revealed the same sOC level between cases and controls except in the European subgroup, where a significantly increased sOC level was observed in PMO patients.The subsequent meta-analysis of all the included studies revealed only a trend of increase in sOC level in PMO cases compared to controls [95%CI: , By sensitivity analysis, we found that the pooled result was significantly influenced by Al-Daghri’s study , in whicThe major result of our current meta-analysis indicated no significant difference in the pooled sOC level between PMO cases and controls, which was consistent with the previous relevant meta-analysis reporting the same sOC level in osteoporotic and healthy populations in 2012 . HoweverHowever, some limitations should be mentioned in this meta-analysis. First, all the included studies were case-control studies due to the design of our research, and the level of evidence was lower than randomized controlled trials (RCTs). Second, significant heterogeneity existed between studies, which could still not be eliminated by subgroup analysis. The heterogeneity between studies might probably come from: 1. variations in the target sOC fragments evaluated by different assay kits; 2. variations in the experimental conditions , 34. TheBased on the current available evidence, there is no significant difference of the pooled sOC level in PMO cases compared to postmenopausal controls. Since OC molecules are quite heterogeneous (different carboxylation status and different fragments) in the circulation and can be influenced by multiple metabolic events, sOC is not a good indicator for the high bone turnover status in PMO unless novel techniques for standardized circulatory sOC evaluation are applied in the future. Despite our rigorous methodology, the level of available evidence can still be improved with further high quality studies.


Gallstone disease (GD) is one of the most common presentations to surgical units worldwide and shares several risk factors with cardiovascular disease (CVD). CVD remains the most common cause of death worldwide and results in considerable economic burden. Recent observational studies have demonstrated an association between GD and CVD with some studies demonstrating a stronger association with cholecystectomy. We present the findings of a meta-analysis assessing the relationship between GD and CVD. A total of fourteen cohort studies with over 1.2 million participants were included. The pooled hazard ratio  for association with GD from a random-effects model is 1.23 (95%CI: 1.16–1.30) for fatal and non-fatal CVD events. The association was present in females and males. Three studies report the relationship between cholecystectomy and CVD with a pooled HR of 1.41 (95%CI: 1.21–1.64) which compares to a HR of 1.30 (95%CI: 1.07–1.58) when cholecystectomy is excluded although confounding may influence this result. Our meta-analysis demonstrates a significant relationship between GD and CVD events which is present in both sexes. Further research is needed to assess the influence of cholecystectomy on this association. Despite significant advances in the prevention and treatment of CVD, the societal and economic burden is considerable and continues to rise4. CVD is preventable and it is cost-effective to promote risk factor modification in those at risk of CVD5. Early identification of at-risk populations may allow these changes to be implemented at a time when they are most effective.Cardiovascular disease (CVD) remains the most common cause of death worldwide8 and patients with gallstones have a higher prevalence of risk factors for cardiovascular disease including obesity and elements of the metabolic syndrome10. The presence of gallstones is associated with atherosclerosis11 and patients with gallstones may be at high risk of progression to symptomatic CVD.Gallstone disease (GD) forms one of the most common presentations to surgical units worldwide22. While recent meta-analyses have demonstrated an association between GD and CVD, there is significant variation in the effect size of contributing studies. No meta-analysis to date has adequately accounted for confounding and the heterogeneity remains unexplained24. Further uncertainty is present over the role of cholecystectomy in modifying the risk of CVD with some authors concluding that cholecystectomy may directly cause CVD24. Analyses of cholecystectomy and CVD have not sufficiently accounted for the presence of confounding variables such as gallstone severity and inflammation. It is likely that these factors account for any apparent association between cholecystectomy and CVD. An additional cohort study examining the relationship between screen-detected GD and CVD has also been published with assessment of the impact of cholecystectomy on CVD22.Several observational studies have been published reporting a link between GD and CVD although some studies have found no differenceWe therefore undertook a systematic review with meta-analysis of longitudinal studies assessing the impact of GD on CVD. We performed additional analyses assessing the impact of cholecystectomy on this association.14 and one publication reported three separate cohort studies as well as a meta-analysis21. Two cohort studies included participants recruited from the same National Health Insurance Research Database (Taiwan) and recruited controls from overlapping inclusion periods18. To prevent bias in our meta-analysis we conducted all analyses twice: first using the larger of the two studies18 and second using the smaller study17. We present results from the first set of analyses and describe any differences from the second set of analyses.The search identified 5497 references of which 984 were duplicates. Searching of bibliographies identified six other potentially relevant references. Following screening of titles and abstracts, 118 articles remained for full text review. After exclusion of 93 articles , a total of 25 citations referring to fourteen cohort studies were included in this systematic review . Two studies reported in the same publication report odds ratio (OR) and are considered in the qualitative analysis. Two studies report relative risk (RR) of which one does not report a sample size and the studies were therefore considered qualitatively. One study does not report numeric data. Overall this review includes over 1,269,137 participants (number of participants not provided in two studies). The quantitative meta-analysis included 1,229,171 participants .Ten of the fourteen studies reported a statistically significant increase in CVD due to GD. Three studies report non-statistically significant findings of which two demonstrate a trend towards an increase in CVD with GD and one reports a trend towards reduced CHD with GD in a female-only cohort. The study reporting no numeric data examined CHD and states: “There was no significant relationship with a history of gallbladder disease”.2 = 76.3%, p = 0.0001).After controlling for confounding variables participants with GD had a 23% increased risk of CVD compared to controls .Seven studies reported CHD as a separate outcome including 843,312 participants . Participants with GD had a 24% increased risk of CHD compared to controls (95% CI = 1.16–1.32). We identified considerable heterogeneity when CHD was considered .Four studies reported cerebrovascular disease as a separate outcome although two studies used controls from the same database. Three studies were pooled by meta-analysis including 458,518 participants . Participants with GD had a 29% increased risk of cerebrovascular disease compared to controls (95% CI = 1.26–1.32). We did not identify heterogeneity when cerebrovascular disease was considered  when both Taiwanese studies using data from the same database were excluded.We performed sensitivity analyses by iteratively excluding one study from the analysis to detect its impact on heterogeneity. Heterogeneity remained substantial for all sensitivity analyses with one study removed. Heterogeneity was noted to fall .We performed a meta-regression which did not detect any significant effect of follow-up, year of publication, quality assessment score, degree of adjustment for confounding variables or variance . In addition, none of the explanatory variables accounted for heterogeneity (pseudo-R2 = 27.8%). Pooled adjusted HR for GD without cholecystectomy compared to controls was 1.30 (95%CI:1.07–1.58) with moderate heterogeneity . The study compared cholecystectomy to GD without cholecystectomy directly found no statistically significant difference . One study also reported mild GD (cholelithiasis or choledocholithiasis excluding definition of severe GD) and severe GD  and reported a stronger association in mild gallstone disease .Three studies report separate data for cholecystectomy compared to controls and for GD without cholecystectomy compared to controls. One study reports a separate analysis of cholecystectomy versus GD without cholecystectomy. Pooled adjusted HR for cholecystectomy compared to controls was 1.41 (95%CI: 1.21–1.64) with insignificant heterogeneity  and found a non-significant trend towards lower risk of CVD in patients undergoing cholecystectomy compared to those with gallstones who did not undergo cholecystectomy.Overall four studies favour a stronger relationship between GD and CVD when cholecystectomy alone is considered and six studies favour a weaker relationship or no difference. No studies report a statistically significant difference between the two groups. Only one studyWe did not detect publication bias through visual inspection of the funnel plot or by Egger’s regression test  statementIn October 2017 we ran searches in MEDLINE, EMBASE, Science Citation Index and Social Sciences Citation Index Expanded and Literatura Latino-americana e do Caribe em Ciências da Saúde (LILACS) from earliest date possible. The search strategies are shown in Supplementary Table Longitudinal cohort studyAssessing the relationship between GD and CVDWe included studies meeting the following criteria:We included cholecystectomy as well as GD without cholecystectomy, we also included screen-detected as well as symptomatic GD. Studies which reported “gallbladder disease” and were thought to represent gallstones rather than rarer gallbladder pathology were considered only in the qualitative analysis. We included fatal and non-fatal CVD events and considered CHD, cerebrovascular disease and peripheral vascular disease (PVD). Each subgroup of CVD was considered separately in subanalyses where possible as well as combined into one single composite outcome of CVD. No relevant non-English language articles were identified. When identified studies were reported in multiple publications we extracted unique data from each publication.39. Prior to data extraction we defined an appropriate follow-up period of at least 10 years to allow for CVD events to occur and a follow-up rate of >90%. We contacted authors to request additional data when required.We extracted the following data from each study: authors, publication year, study population, baseline characteristics, sample size, study design, country, ascertainment of exposure, detection of CVD events, definition of CVD, follow-up duration, confounding variable adjustment, rate of cholecystectomy and adjusted effect measures  with 95% confidence interval. We assessed each study according to the Newcastle-Ottawa Quality Assessment Scale (NOS)40. We used fully adjusted (multivariable) effect measures from each study when available. We did not transform each unique effect measures into one single effect measure and as such studies reporting HR, RR or OR are considered in three distinct groups. All effect measures were log-transformed for purposes of data analysis prior to back-transformation for presentation of results.We assimilated data using DerSimonian-Laird random effects meta-analysisMale-only participantsFemale-only participantsDiabetic participantsNon-diabetic participantsScreen-detected GDSymptomatic GDGallstones (not including cholecystectomy) versus controlsCholecystectomy only versus controlsSubgroup analyses were performed for the following groups:41. For the mixed effects model and meta-regression we included length of follow-up, year of publication, degree of adjustment for confounding variables, variance and NOS score.Discrete subgroups were compared using a Wald-type test with p value significance set at <0.01. We assessed influences of moderator variables by meta-regression using a mixed effects model and the Knapp and Hartung adjustment2 statistic43. We assessed for the presence of publication bias by visual inspection of funnel plots and Egger’s regression test with p value significance set at <0.144.We assessed for presence of heterogeneity using Cochran’s Q test with p value significance set at <0.1 and quantified heterogeneity with the I46. All data is available in original manuscripts.All analyses were undertaken in R version 3.3.1 using the “metafor” packageSupplementary Information