Micro RNAs (miRNAs) are small RNA fragments that naturally exist in the human body. Through various physiological mechanisms, miRNAs can generate different functions for regulating RNA protein levels and balancing abnormalities. Abnormal miRNA expression has been reported to be highly related to several diseases and cancers. Single-nucleotide polymorphisms (SNPs) in miRNAs have been reported to increase patient susceptibility and affect patient prognosis and survival. We adopted a case-control research design to verify the relationship between miRNAs and hepatocellular carcinoma.A total of 525 subjects, including 377 controls and 188 hepatocellular carcinoma patients, were selected. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and real-time PCR were used to analyze miRNA146a (rs2910164), miRNA149 (rs2292832), miRNA196 (rs11614913), and miRNA499 (rs3746444) genetic polymorphisms between the control group and the case group. The results indicate that people who carry the rs3746444 CT or CC genotypes may have a significantly increased susceptibility to hepatocellular carcinoma . In addition, when combined with environmental risk factors, such as smoking and alcohol consumption, interaction effects were observed between gene polymorphisms and environmental factors .These results suggest that a significant association exists between miRNA499 SNPs and hepatocellular carcinoma. Gene-environment interactions of miRNA499 polymorphisms, smoking, and alcohol consumption might alter hepatocellular carcinoma susceptibility. Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide and the second leading cause of cancer-related death in Taiwan. In addition to common risk factors such as hepatitis B and hepatitis C, recent studies have shown that gene mutations may influence the inflammation mechanism that contributes to cell differentiation, proliferation, and apoptosis and connects several diseases and cancers Micro RNAs (miRNAs) are small RNA fragments typically composed of only 20–22 nucleotides. MiRNAs can target specific mRNAs and negatively regulate their translational efficiency and stability to control the downstream cellular process including proliferation, differentiation, and survival Single-nucleotide polymorphisms (SNPs) are variations of DNA sequences that occur when nucleotides  are changed by at least 1% in certain populations. Previous epidemiologic studies have shown that genetic variations in miRNAs are associated with various diseases and cancers Based on the aforementioned reasons, we adopted a case-control research design and 4 pre-miRNA SNPs associated with cancer, namely, has-mir-146a (rs2910164), has-mir-149 (rs2292832), has-mir-196a2 (rs11614913), and has-mir-499 (rs3746444) This study recruited 188 HCC patients at the Chung Shan Medical University Hospital, Taiwan. The diagnosis of HCC was made according to the criteria specified in the national guidelines for HCC. During the same study period, 337 ethnic group-matched individuals were enrolled as the controls that entered the physical examination at the same hospital. These control groups had neither self-reported history of cancer of any sites. As for cases and controls, exposure information, including tobacco use (smoker vs. non-smoker) and alcohol consumption , were all obtained from questionnaires. We also collected the laboratory status such as α-Fetoprotein, AST, ALT and AST/ALT ratio for the further analysis. Before commencing the study, approval was obtained from the Institutional Review Board of Chung Shan Medical University Hospital and informed written consent was obtained from each individual.We collected the whole blood samples from healthy controls and hepatocellular carcinoma patients with tubes containing EDTA, after centrifuged and stored at −20°C. The venous blood from each subject was drawn into Vacutainer tubes containing EDTA and stored at 4°C. Genomic DNA was extracted by QIAamp DNA blood mini kits  according to the manufacture’s instructions and the DNA was dissolved in TE buffer  and then quantitated by a measurement of OD260. Final DNA preparation was stored at −20°C and used as templates for the following experiments 5′- CATGGGTTGTGTCAGTGTCAGAGCT-3′ and the reverse primer 5′- TGCCTTC TGTCTCCAGTCTTCCAA-3′; the has-mir-196a2 rs 11614913 genotype were used forward primer 5′- CCCCTTCCCTTCTCCTCCAGATA-3′ and reverse primer 5′- CGAAAACCGACTGATGTAACTCCG-3′ and the has-mir-499 rs3746444 genotype were used forward primer 5′-CAAAGTCTTCACTTCCCTGCCA-3′ and reverse primer 5′-GATGTTTAACTCCTCTCCACGTGATC-3′. PCR was performed with total 10 µl volume with 100 ng DNA template, 1.0 µl of 10X PCR buffer , 0.25 U of Taq DNA polymerase (Invitrogen), 0.2 mM dNTPs  and 200 nM of each primer . The PCR conditions were 5 min at 94°C followed by 35 cycles of 30 sec at 94°C, 30 sec at 58°C for hsa-mir-146a rs2910164, 30 sec at 63°C for has-mir-196a2 rs11614913, and 30 sec at 67°C for hsa-mir-499 rs3746444, and 1 min at 72°C, and final step at 72°C for 10 min to allow a complete extension of all PCR fragments. PCR products of hsa-mir-146a rs2910164 gene polymorphism were subjected SacI to enzymatic digestion by incubation for 4 hr at 37°C and then submitted to electrophoresis in 2% agarose gels; MspI for hsa-mir-196a2 rs2910164 and BclI for hsa-mir-499 rs3746444.The SNP of miRNAs gene rs2910164, rs11614913, and rs3746444 polymorphisms were determined by PCR-RFLP assay MiRNA149 rs2292832 gene polymorphism was assessed using an ABI StepOne™ Real-Time PCR System (Applied Biosystems), SDS v3.0 software (Applied Biosystems), and the TaqMan assay. The final volume for each reaction mixture was 5 µL, containing 2.5 µL TaqMan Genotyping Master Mix, 0.125 µL TaqMan probes mix, and 10 ng genomic DNA. The reaction conditions included an initial denaturation step at 95°C for 10 min followed by 40 cycles at 95°C for 15 sec and 60°C for 1 min. For each assay, appropriate controls (nontemplate and known genotype) were included in each typing run to monitor reagent contamination and as a quality control. To validate results from real-time PCR, around 5% of assays were repeated and several cases of each genotype were confirmed by the DNA sequence analysis p value of less than 0.05 was considered significant. The data were analyzed on SPSS 12.0 statistical software.Hardy–Weinberg equilibrium was assessed using a chi-square goodness-of-fit test for biallelic markers. The distributions of demographic characteristics and genotype frequencies between cases and controls in different genotypes were analyzed by Chi-square test, and Fisher’s exact test were using at small sample size was present in some categories of variables. The Student’s T-test was using to estimate laboratory status between the two groups. The odds ratios (ORs) and their 95% confidence intervals (CIs) of the association between genotype frequencies and hepatocellular carcinoma were estimated by multiple logistic regression models, also controlling for covariates. The p = 0.03).The demographic characteristics of the 2 groups, including age, sex, smoking status, and alcohol consumption, are shown in 2 value: 1.92), miRNA196 rs11614913 , and miRNA499 rs3746444  were in Hardy-Weinberg equilibrium, respectively, except for miRNA149 rs2292832 . By using a multiple logistic regression model to estimate the adjusted odds ratio (AOR), we observed a significantly higher risk for people who carried the CT  or CC  genotypes of miRNA499 (rs3746444). Because of the insufficient size of the CC group, the CT and CC genotype samples were combined, producing similar results . In addition, no statistical significance was observed in the other 3 gene polymorphisms . Similarly, a significantly higher risk for people who carried the CT  or CC  genotypes of miRNA499 (rs3746444) was observed in the HBV-infection group .We associated laboratory status with gene polymorphisms, including α-fetoprotein (ng/mL), AST (IU/L), ALT (IU/L), and AST/ALT ratios, as shown in Furthermore, we considered the environmental risk factors of hepatocellular carcinoma such as smoking and alcohol consumption. The risk factors were then combined with miRNA gene polymorphisms and examined to determine whether interactions existed between the 2 factors. The relationship between genotype and smoking status indicated that people who carried rs3746444 CT or CC genotypes and non-smoking habits had an increased odds ratio , and those with CT or CC genotypes and smoking habits had the highest odds ratio  . The resThe results of this investigation indicate that miRNA499 rs3746444 is highly associated with hepatocellular carcinoma. Regarding genotype distribution and environmental risk factors, such as smoking and alcohol consumption, miRNA499 plays a crucial role in hepatocellular carcinoma.MiRNA499 rs3746444 has been discussed in relation to numerous cancers. A meta-analysis of 7188 patients with several cancers compared with 8548 control participants reported that a significant association exists between miR499 rs3746444 polymorphisms and an increased risk of cancer Previous studies have shown that miRNA146 can be used as a biomarker and therapeutic target for peripartum cardiomyopathy A slight difference exists in distant tumor metastasis caused by miRNA149 rs2292832 gene polymorphisms (Table S2 in p = 0.152). Previous studies have shown that miRNA196 is associated with transcription factors and affects cancer development and progression p<0.001) and increase the OR for developing hepatocellular carcinoma. This may be because the various functions of miRNAs not only influence tumor differentiation and proliferation, but also promote inflammation and apoptosis.In this study, miRNA196 exhibited a slight association with hepatocellular carcinoma after patients’ alcohol consumption was considered. However, the test results for an interaction between gene polymorphisms and cancer risk factors were not statistically significant Click here for additional data file.


HOXA1 and HOXB1 have been strongly posed as candidate genes for autism spectrum disorders (ASD) given their important role in the development of hindbrain. The A218G (rs10951154) in HOXA1 and the insertion variant in HOXB1  were of special interest for ASD but with inconclusive results. Thus, we conducted a meta-analysis integrating case-control and transmission/disequilibrium test (TDT) studies to clearly discern the effect of these two variants in ASD.Pheterogeneity = 0.029) and 1.14 , respectively. No significant association was also identified between these two variants and ASD risk in stratified analysis. Further, cumulative meta-analysis in chronologic order showed the inclination toward null-significant association for both variants with continual adding studies. Additionally, although the between-study heterogeneity regarding the A218G is not explained by study design, ethnicity, and sample size, the sensitive analysis indicated the stability of the result.Multiple electronic databases were searched to identify studies assessing the A218G and/or nINS/INS variant in ASD. Data from case-control and TDT studies were analyzed in an allelic model using the Catmap software. A total of 10 and 7 reports were found to be eligible for meta-analyses of A218G and nINS/INS variant, respectively. In overall meta-analysis, the pooled OR for the 218G allele and the INS allele was 0.97 (95% CI = 0.76-1.25, HOXA1 A218G and HOXB1 nINS/INS variants may not contribute significantly to ASD risk.This meta-analysis suggests the  Autism spectrum disorders (ASD) comprises a pervasive group of neurodevelopmental disorders that share common features of impaired social interaction, deviance in language development, and stereotyped behaviors or narrow range of interests HOXA1 and HOXB1, located on chromosomes 7p and 17q, are two paralogous genes in the HOX gene family of homeobox transcription factors and critically involved in the developing hindbrain during neural tube formation HOXA1 and HOXB1 function for their proper development. Similar morphologic deficits in brain have been described in mice knockout Hoxa1 and Hoxb1Hoxa1 and Hoxb1 are highly expressed during an early period of development overlapping with the window of maximal prenatal sensitivity in rodent model HOXA1 and HOXB1 as potential candidate genes for ASD. Ingram and his collaborators firstly described variants in HOX genes for genetic susceptibility of ASD, including an A to G substitution at base 218  of HOXA1 and a 9-base insertion variant  of HOXB1HOXA1 changes a histidine (H) to arginine (R) at position 73 (H73R) and disrupted a string of histidine repeats in exon 1, while the 9-base insertion c.82insACAGCGCCC  in exon 1 of HOXB1 introduces into the amino acid sequence the tripeptide histidine-serine-alanine (H-S-A) HOXA in 57 autistic probands and 119 unrelated adults as controls, indicating that the G allele carriers might be susceptible to ASD HOXA1, HOXB1, and gender was also found to be associated with increased risk of ASD HOXA1 and HOXB1 in ASD, but the results were inconsistent. Inversely, Conciatori et al. pointed out the A allele, not the G allele, was associated with ASD risk in both case-control and family-based association analyses HOXA1 and HOXB1 genetic variants with ASD risk.http://www.prisma-statement.org).To ensure the rigour of this current meta-analysis, we designed and reported it according to the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) HOXA1 and insertion variant in HOXB1 associated with ASD in case-control study or TDT study. The search strategy was based on combinations of the keywords ‘HOXA1, homeobox A1, HOXB1 or homeobox B1’, ‘polymorphism or variant’ and ‘autism or autistic disorder’ without language restrictions. To expand the coverage of our searches, we further performed searches in Chinese Biomedical (CBM) database using the above searching strategy. References of the retrieved articles were also scanned. Reviews, comments, and letters were also checked for additional studies.We searched PubMed, EMBASE, and ISI Web of Science databases for published articles up to August 2010, which had investigated at least one of A218G in HOXA1 and insertion variant in HOXB1; (3) presentation of data necessary for calculating odds ratios (ORs); (4) clear definition of ASD. Animal studies, reviews, simply commentaries, case reports and unpublished reports were excluded. Study overlapping with other studies should be eliminated, and the report with complete design or larger sample size was finally selected.The inclusion criteria had to be fulfilled: (1) either case-control or TDT study design; (2) data on any or both polymorphisms of A218G in All data were extracted independently by two reviewers (R–R. Song & L. Zou). The following information was extracted from the eligible studies: first author's name, year of publication, ethnicity, design type of study, and diagnostic criteria for ASD. Counts of alleles in case and control group in case-control studies and numbers of transmitted alleles from heterozygous parents to affected offspring in family-based studies were extracted or calculated from published data in the included studies.χ2- based Cochran's Q statistic test was employed to test between-study heterogeneity, and heterogeneity was considered significant when P<0.1 for Q statistic. Data from the studies were combined by the random-effects model when heterogeneity was significantly present; otherwise, fixed-effects model was applied. For the synthesis of case-control and TDT studies, the method described by Kazeem et al. was used http://www.r-project.orgP values are two-tailed with a significant level at 0.05. All statistical analyses were carried out in Catmap software V1.6.Data from case-control studies were summarized in two-by-two tables, and data from TDT studies were summarized in two-by-one tables. ORs and their 95% confidence intervals (CIs) and standard errors (SEs) were calculated for individual study based on the allele data using the method as described by Kazeem et al. HOXA1, and 7 reports including 113 cases, 184 controls and 570 families were relevant to insertion variant in HOXB1. χ2 = 22.83, Pheterogeneity = 0.029); thus, the random-effects model was employed. Pooled OR and 95% CI were calculated for the 218G allele versus the 218A allele. However, no significant association was found between the allelic variant and ASD risk . For HOXB1 nINS/INS variant, no evidence of heterogeneity was presented among the 3 case-control and 5 TDT studies from 7 reports . In the fixed-effects model, this insertion variant was not significantly associated with risk of ASD . There was no indication of heterogeneity for HOXB1 variant . In TDT studies, the heterogeneity test for A218G showed positive results . In the random-effect model, this variant presented no association with ASD risk . No sign of heterogeneity was detected for HOXB1 insertion variant , and no significant association between ASD and this variant was found in the fixed-effects model .The stratified analysis was firstly performed by study design. In case-control studies, evidence of between-study heterogeneity was found for χ2 = 10.10, Pheterogeneity = 0.039) but not in the small-sample size subgroup including 3 case-control studies and 5 TDT studies . No significant association between A218G and ASD risk was found in the small- or large-sample size subgroup . For HOXB1 insertion variant, no significant associations were observed in both the large-sample-size subgroup including 3 TDT studies  and the small-sample size subgroup including 3 case-control studies and 2 TDT studies .The data were further stratified by sample size into 2 subgroups, the large-sample-size subgroup (number of cases in case-control study or number of family in family-based study >80) and the small- or moderate-sample-size subgroup (number of cases in case-control study or number of family in family-based study ≤80). Heterogeneity was detected for A218G in the large-sample-size subgroup including 5 TDT studies (Pheterogeneity = 0.002) and the latter was 1.17 . The pooled OR could not be appraised in Blacks, Yellows, and Indians because of limited numbers of studies conducted in these populations.In terms of ethnicity, the data were stratified into Whites, Blacks, Yellows, and Indians. No statistically significant finding was seen in Whites for A218G or nINS/INS variant. The pooled OR for the former was 0.92 , but the effect of 218G still lacked of significance .Given the significant between-study heterogeneity for ASD risk . AdditioHOXA1 A218G in the random-effects model and the HOXB1 nINS/INS in fixed-effects model. The effect of A218G and nINS/INS both tended to be null significant association over time. Moreover, the 95% CIs became increasingly narrower with increasing data, suggesting that the precision of the estimates was progressively enhanced by continual adding more studies.Cumulative meta-analyses of these two variants were also conducted via assortment of studies in chronologic order. P = 0.564 and 0.591, respectively).As reflected by funnel plots  and EggeHOXA1 and 113 cases, 184 controls, and 570 families concerning the insertion variant in HOXB1. Both the A218G and nINS/INS variants showed no significant association with risk of ASD. This current meta-analysis, to the best of our knowledge, firstly integrated the case-control and TDT studies to reflect the precision effect of HOXA1 and HOXB1 variants in ASD risk.This current meta-analysis included 113 cases, 184 controls, and 958 families concerning the A218G in HOXA1 is a critical member of HOX gene family involved in the development of hindbrain HOXA1 was of special interest in ASD. In the HOXA1 gene, the A218G variant causing a histidine to arginine has been shown to disrupt the string of histidine repeats, which was believed to be the binding site of other proteins HOXB1, in conjunction with HOXA1, partly determined the placement of hindbrain segments in the proper location along the anterior-posterior axis during embryo development HOXB1 gene Neuregulin 1 polymorphisms in schizophrenia HOXA1 or HOXB1 variant, indicating the statistical consistency between these two types of studies. When the case-control studies were combined with family studies, statistical power was enhanced from enlargement of sample size, and the ORs and P values stably showed the null-significant associations. This meta-analysis integrating case-control with family-based studies therefore provides a straightforward mean to increase the ability to clarify the conflicted results of genetic studies.Integrating case-control and family based studies is an important aspect of genetic analysis, and combined estimates can provide an overall picture of the effect size attributable to genetic polymorphism. An illustrative application has been made by Gong et al. to HOXB1 and HOXA1 genes have synergized in patterning hindbrain structures, there was a hypothesis that gene-gene interaction between HOXA1 and HOXB1 variants might be involved in ASD. However, interaction of these two variants could not be appraisal in this meta-analysis because of a lack of special data.Despite the clear strength of this study integrating case-control and family studies, some limitations merit serious consideration. Some heterogeneous natures of studies, including mixed population samples, variant age range of controls in case-control design, multiplex or simplex trios utilized in family studies, possibly have effect on the current result. However, due to lack of detail data or limited small number of some studies, we were unable to perform further analysis. Given the HOXA1 and HOXB1 variants with ASD, and suggest null association of the A218G and nINS/INS variants with ASD risk. Further analysis should be imposed in possible interaction effect between HOXA1 and HOXB1 genetic polymorphisms in modulation of ASD risk.In conclusion, this meta-analysis helped for strongly clarifying the discrepancies of genetics studies into associations of Table S1PRISMA Checklist for this meta-analysis.(DOC)Click here for additional data file.Figure S1The funnel plots of the HOXA1 A218G variant for ASD. Egger's test: P = 0.564.(TIF)Click here for additional data file.Figure S2The funnel plots of the HOXB1 nINS/INS variant for ASD. Egger's test: P = 0.591.(TIF)Click here for additional data file.


Numerous studies have investigated the relationship between apolipoprotein (Apo) E gene polymorphisms and gallbladder stone disease (GSD) across ethnic populations; however, the results are often inconsistent. This meta-analysis aims to comprehensively evaluate the influence of a common ε2/ε3/ε4 polymorphism in Apo E gene on the risk of gallbladder stone disease.Data were analyzed using the RevMan software (V5.1) and a random-effects model was applied irrespective of between-study heterogeneity. Publication bias was weighed using the fail-safe number.2I = 13%), and comparison of alleles ε4 with ε3 yielded a 25% increased risk . Subgroup analysis by study design indicated that the magnitude of association in hospital-based studies was largely significantly strengthened for ε4 allelic model . Subgroup analysis by age of controls indicated a remarkably significant elevation in the magnitude of association in age >50 subgroups in ε4 allelic model . Moreover, subgroup analysis by cases gender indicated a reduction in the magnitude of association in male<30% studies for E2/2 genotypic model .There were 17 study populations totaling 1773 cases and 2751 controls for ε2/ε3/ε4 polymorphism of Apo E gene. Overall comparison of alleles ε2 with ε3 in all study populations yielded a 16% decreased risk for GSD (95% confidence interval [95% CI]: 0.68–1.05; P = 0.31; Our results reveal that Apo E gene ε4 allele is a risk factor of gallbladder stone disease, especially in elder people and Chinese population. Gallbladder stone disease (GSD) is one of the most common digestive disorders worldwide, especially in western population One of the candidate lithogenic-genes had been studied is apolipoprotein (Apo) E. It is a ligand for the low density lipoprotein family of receptors and plays a pivotal role in cholesterol metabolism http://www.ncbi.nlm.nih.gov/pubmed/), Embase (http://www.embase.com), ISI Wed of Knowledge (http://isiknowledge.com), Wanfang (http://www.wanfangdata.com.cn) and China Biological Medicine (CBM) (http://cbm.imicams.ac.cn) with the last update as of June 2012. The keywords used for search were ‘gallbladder stone disease’ and ‘Apolipoprotein E or Apo E’ combined with ‘gene or variants or polymorphism or alleles’, all of which were MeSH terms . The ‘related articles’ option in MEDLINE, as well as reference lists of all retrieved studies, were checked to search for other relevant publications that were not initially identified. As a prerequisite, only these published in English or Chinese language were identified, and studies in human subjects. In addition, the full text of the retrieved articles was scrutinized to make sure the data of interest were included. If two or more studies shared the same cases or control subjects, the one with small sample size was abandoned. If more than one geographical or ethnic population were included in one article, each population was considered separately.Publication were searched via public database PubMed  and 95% confidence interval (95% CI).Gallbladder stone disease was diagnosed by operation or ultrasound. In some studies, the cholesterol content in gallstone was analyzed. At the same time, some GSD with T2DM, hypertension, pregnancy, or underwent bariatric surgery were also included.Studies were excluded if they were prospective studies; published in minor language; or published abstracts from meeting.Two authors (P. Xue and WQ. Niu) independently drew the following information from all qualified studies: first author’s last name, publication date, population ethnicity, methods to diagnosis of gallstone, study design, methods of genotyping, the distribution of alleles and the genotype in cases and controls. Information such as cases and controls’ age, BMI, and serum concentration of TC, TG, LDL, HDL were also collected. The units of measurements used in this study were transformed into the standard measurement units.The associations between genotypes/alleles of Apo E polymorphism with GSD were evaluated by using the software Review Manager (V5.1) for windows. In this meta-analysis, we used the random-effects model with the method of DerSimonian & Laird to bring the individual effect-size estimates together. The estimate of heterogeneity was taken from the Mantel-Haenszel model 2I statistic, which was documented for the percentage of the observed between-study variability due to heterogeneity rather than chance with the ranges of 0 to 100%  Heterogeneity was assessed by the fs) with the significance set at 0.05 for each meta-comparison. Specifically, if the calculated Nfs value was smaller than the number of observed studies, than the meta-analysis results might run the risk of having publication bias. We calculated the Nfs0.05 according to the formula Nfs0.05 =  (ΣZ/1.64)2–k, where k is the number of included articles We assessed publication bias using the fail-safe number  at the first time. According to our inclusion/exclusion criteria, we removed 5 of them for the following reasons: three papers shared same study group, one review, one without data of genotype distribution. Sixteen qualified studies .  . No sign = 13%). .2I statistic indicated the large between-study heterogeneity for all comparisons except for that of E4/4 with E3/3 (2I = 0) and recessive models (2I = 0).  . There wI2 = 0). .Han population, after restricting analyses to Han populations, there was no material change in OR for both polymorphisms under study (data not shown).Moreover, since most of studies were conducted in Chinese Considering the fact that study design, samples age, gender distribution or ethnic differences might bias the overall association, we conducted separate analysis according to these factors.In view of study design, the magnitude of association in population-based studies was weakened for ε2 allelic model , ε4 allelic model , E3/4 genotypic model  and E4/4 genotypic model . Meanwhile, the magnitude of association in hospital-based studies was largely significantly strengthened for ε4 allelic model , E3/4 genotypic model , E4/4 genotypic model , ε2 recessive model  and ε4 dominant model . Nearly no change in ORs was observed in other subgroup analysis for study design.The age of onset of GSD differed from different studies. Since GSD prevalence increases with age, to avoid the miss-classification, we choose age of 50 as the cut-off in controls to divide the studies into two subgroups. A remarkable significantly strengthen in the magnitude of association in age >50 subgroups in ε4 allelic model , E3/4 genotypic model  and ε4 dominant model  could be observed. No significant change in ε2 models or other subgroups was present.The ratio of GSD in female and male is about 2∶1, thus we choose the 30% gender percentage  as the cut-off to divide populations into two subgroups, considering that the number of each subgroup is almost equal at the same time. We found that the magnitude of association in male <30% studies was strengthened for E2/2 genotypic model  and the magnitude of association in male >30% studies was strengthened for E4/4 genotypic model . This suggests that ε2 may provide protection against GSD in women, and on contrary, ε4 is a risk gene promoting GSD. Although the prior viewpoint has been proposed by Niemi et al in 1999 In order to control for the difference of ethnicity, we separated the studies in Chinese and non-Chinese. The magnitude of association in Chinese studies was significantly strengthened for both ε4 allelic model  and ε4 dominant model . There was no significant change in other subgroups.fs) was shown in The result of publication bias assessed by the fail-safe number  to evaluate the association of Apo E gene polymorphisms with GSD. To the authors’ knowledge, this is the first meta-analysis investigating the association between Apo E ε2/ε3/ε4 polymorphisms and GSD. Although some statistical bias could not be eliminated, this meta-analysis suggests that the Apo E ε4 allele appears to be associated with an increased risk of GSD and also appears to bedominant.Previous researches have shown that the presence of the ε4 allele of Apo E is strongly associated with the risk of atherosclerosis Actually, in the newest review by Laura M. Stinton and Eldon A. Shaffer The prevalence of gallstone disease increases with age, escalating markedly after middle age, becoming 4–10 times more likely in older individuals By contrast, the conclusion of ε2 allele is equivocal. It seems that ε2 allele can provide a protection against GSD in women. Nimei Last but not least, subgroup analysis by study design showed that hospital-based studies yielded a more significant association signals than population-based studies. Generally, in population-based studies, it is not clear if the people in controls had other diseases which could exert a confounding effect on the true association. Only 4 publications were population-based study, it is still too little for the meta-analysis. Thus, the results on study design should be more cautious for interpretation.Despite the clear strength of our study including large sample sizes, some limitations merit serious consideration. First, all included studies were cross-sectional design, which precludes further comments on cause-effect relationship Taken together, in this meta-analysis, we ascertained the role of Apo ε4 allele on the risk factor of GSD. Although the publication bias was maximally avoided, presence of between-study heterogeneity could not be fully explained by our subgroup analysis. Nowadays further analyses are warranted to investigate Apo E gene adjacent markers in a wider context, future studies on Apo E and GSD should concentrate on gene-environment interactions.Table S1(XLS)Click here for additional data file.


CA9 gene polymorphisms on the susceptibility of developing urothelial cell carcinoma (UCC) and the clinicopathological status.Carbonic anhydrase 9 (CA9) is reportedly overexpressed in several types of carcinomas and is generally considered a marker of malignancy. The current study explored the effect of CA9 gene were assessed by a real-time PCR with the TaqMan assay. After adjusting for other co-variants, the individuals carrying at least one A allele at CA9 rs1048638 had a 2.303-fold risk of developing UCC than did wild-type (CC) carriers. Furthermore, UCC patients who carried at least one A allele at rs1048638 had a higher invasive stage risk (p< 0.05) than did patients carrying the wild-type allele. Moreover, among the UCC patients with smoker, people with at least one A allele of CA9 polymorphisms (rs1048638) had a 4.75-fold (95% CI = 1.204–18.746) increased risk of invasive cancer.A total of 442 participants, including 221 healthy people and 221 patients with UCC, were recruited for this study. Four single-nucleotide polymorphisms (SNPs) of the CA9 might contribute to the prediction of susceptibility to and pathological development of UCC. This is the first study to provide insight into risk factors associated with CA9 variants in carcinogenesis of UCC in Taiwan.The rs1048638 polymorphic genotypes of  Urothelium covers the epithelial lining of the urinary tract from the renal calyces to the bladder. Any neoplastic lesion grew from the urothelium has similar behavior2 and HCO3−. Carbonic anhydrase 9 (CA9) is located on chromosome 9p12–13, which comprises 11 exons and encodes for the 459-amino-acidprotein. CA9 helps to keep a normal pH in tumor cells in a hypoxic microenvironment and allow tumor cell proliferation Carbonic anhydrase (CA), a family of zinc metalloenzymes, can efficiently catalyze the reversible processes of hydration-dehydration of COWe recruited 221 patients  at Taichung Veterans General Hospital in Taichung, Taiwan. Patients were enrolled as a case group in 2011–2012. All patients have pathology proved urothelial cell carcinoma of upper urinary tract or bladder. Meanwhile, during the same study period, 221 age- and gender-matched individuals were enrolled as the controls that entered the physical examination at the same hospital. These control groups had neither self-reported history of cancer of any sites. Personal information and characteristics collected from the study subjects using interviewer-administered questionnaires contained questions involving demographic characteristics and the status of cigarette smoking. Before commencement of this study, approval was obtained from the Institutional Review Board (IRB) of Taichung Veterans General Hospital, and informed written consent to participate in the study was obtained from each person (IRB No. CF11094). For both cases and controls, we used a questionnaire to obtain information on patient exposure to tobacco consumption. Medical information for the cases was obtained from their medical records, and included TNM clinical staging, lymph node involvement, and histologic grade. UCC patients were clinically staged at the time of diagnosis according to the TNM staging system of the American Joint Committee on Cancer (AJCC) Staging Manual (7th ed.) Tumors are classified as superficial tumor  and invasive tumor . Metastasis into lymph nodes was detected in 11 cases (6.1%) and one patient has distal metastasis (0.6%). Whole-blood specimens collected from controls and UCC patients were placed in tubes containing ethylenediaminetetraacetic acid (EDTA), immediately centrifuged, and stored at –80°C Genomic DNA was extracted using QIAamp DNA blood mini kits  following the manufacturer’s instructions. We dissolved DNA in TE buffer  and then quantified it by measuring the OD260. The final preparation was stored at –20°C and was used to act as templates for the polymerase chain reaction (PCR) CA9 +201 (rs2071676), +1081 (rs3829078) and +1584 (rs1048638) allelic polymorphisms were assessed with an ABI StepOne™ Real-Time PCR System , and analyzed with SDS vers. 3.0 software (Applied Biosystems) using the TaqMan assay Allelic discrimination of the p values were < 0.05. The Mann-Whitney U-test and Fisher’s exact test were used to compare differences in distributions of patient demographic characteristics between the cancer-free (control) and UCC groups. The adjusted odds ratios (AORs) and 95% confidence intervals (CIs) of the association between genotype frequencies and risk plus clinicopathological characteristics were estimated using multiple logistic regression models, after controlling for other covariates. We analyzed all data with Statistical Analytic System  software  for Windows.Differences between the 2 groups were considered significant if CA9 rs2071676 , rs3829078 , rs1048638 , and 376del393  were in Hardy-Weinberg equilibrium, respectively. Distribution of CA9 genotypes in The distributions of demographic characteristics showed no significant differences in age, gender and tobacco consumption between UCC patients and controls . The stuStudy subjects carrying the C/A genotype of the rs1048638 polymorphism had a significantly increased UCC risk of 2.217  polymorphism had a significantly increased UCC risk of 2.217 (95% CI = 1.169–4.207). Compared to C/C genotype of the rs1048638, C/A+A/A genotype demonstrated higher UCC risk of 2.303 (95%CI = 1.227–4.320). To consider the tumor invasiveness, patients with C/A+A/A genotype of the rs1048638 polymorphism demonstrated a significant higher invasive caner (pT2-T4) risk of 2.259 (95% CI = 1.136–4.490).In the current study, we try to find the association between CA9 is abundant in cancers but not in most benign tissues. It helps tumor cells to survive and proliferate in a hypoxic condition CA9 reflects significant changes in tumor biology and decreased CA9 concentration is independently associated with poor survival in advanced renal cell carcinoma. CA9 can be used to predict clinical outcome in such high risk kidney cancer patients who need adjuvant immunotherapy and CA9 targeted therapies Alternation of genome can change the cellular phenotype to evolve from a preneoplastic stage into cancer The study still has some limitations. Firstly, the sample size was small; the statistical significance should be interpreted carefully. Secondly, patients cannot provide detail history of Chinese herbs usage or aromatic amines exposure that might cause bias. We observed the CA9 SNPs affected the clinical behavior of UCC but how this SNP influenced the CAIX expression was still unknown.In conclusion, this is the first report to study the association between CA9 SNP and UCC risk. The patients carrying the C/A genotype of the rs1048638 of the CA9 gene polymorphism had a significantly increased UCC risk and invasive cancer risk. The synergistic effect of smoking and CA9 gene polymorphisms on the risk of invasive UCC is also demonstrated. Large-scale studies may be required to verify the association between the SNP of hypoxia related gene such as CA9 and UCC risk.


Preeclampsia, characterized by hypertension and proteinuria, is a multifactorial disease caused by complex interactions between environmental and genetic factors. A recent genome-wide association study of blood pressure reported an association between hypertension and rs11646213. This study evaluated the association between preeclampsia and rs11646213.A total of 454 cases and 460 controls were recruited to participate in this study. The single nucleotide polymorphism (SNP) rs11646213 was genotyped by polymerase chain reaction (PCR) and direct sequencing.The allele frequency of rs11646213 was significantly different between the preeclampsia and control groups . Differences were particularly significant in the severe preeclampsia subgroup  and the early-onset preeclampsia subgroup . Genotyping analysis showed that the T allele of rs11646213 could confer a risk for preeclampsia, severe preeclampsia and early-onset preeclampsia.CDH13 gene is associated with preeclampsia in Han Chinese women.Rs11646213 upstream of the  Preeclampsia affects approximately 2.5% to 3% of all pregnancies and is a leading cause of maternal morbidity and mortality in developing countries CDH13) gene is associated with hypertension in Europeans Epidemiological studies have shown that women with chronic hypertension are at an increased risk of developing preeclampsia and that a family history of chronic hypertension is more common in preeclamptic patients compared with controls Thus, we proposed that SNP rs11646213 might be a genetic risk factor for preeclampsia and investigated the association between preeclampsia and rs11646213 in a cohort of Han Chinese women.The study was approved by the Institutional Review Board of the Provincial Hospital Affiliated to Shandong University. And written informed consent was obtained from each participant.A total of 914 Han Chinese pregnant women who visited Provincial Hospital Affiliated to Shandong University were enrolled from December 2009 to November 2011. Controls were normotensive women with no antenatal medical or obstetric complications, presenting normal fetal growth at ≥37weeks of gestation (birth weight between the 10th and the 90th percentile), and with a regular postpartum course. Preeclampsia was defined as gestational hypertension with proteinuria  after the 20th week of pregnancy in a previously normotensive and non-proteinuric woman Blood pressure was measured in the right arm after a 5-minute period of rest. The following clinical data were collected: maternal age, parity, gestational age at birth, fetal weight and prepregnancy body mass index (pre-BMI). Participants were excluded for the following reasons: a previous renal, autoimmune, metabolic or cardiovascular disease, chronic hypertension, lost to follow-up and multiple pregnancies. Women with gestational hypertension, which is defined as elevated blood pressure without proteinuria, were also excluded from the study.Genomic DNA was extracted by QIAamp DNA mini kit . Polymerase chain reaction (PCR) conditions were as followings: 95°C for 10 minutes, followed by 34 cycles at 95°C for 30 seconds, 58°C for 30 seconds, 72°C for 45 seconds, and a final step at 72°C for 10 minutes.The primer sequences were:5′-AAGGAGGGAAGTGTTGACC-3′;Forward: 5′-GTCTGGCTTTCATTTCTGG -3′.Reverse: The products were analyzed by agarose gel electrophoresis and then were sequenced by an automated sequencer .General characteristics of the cases and controls were expressed as mean±SD. Linkage disequilibrium (LD) was assessed using Haploview software . The chi-square test was performed to compare allele frequency. Genetic models were divided into additive (+/+ vs. +/− vs. −/−), dominant (+/+ plus +/− vs. −/−) and recessive (+/+ vs. +/− plus −/−) groups and analyzed by one-way ANOVA. Unconditional logistic regression analysis was used for parity, pre-BMI and maternal age adjustments using SPSS statistical software . P<0.05 was considered statistically significant.The clinical characteristics of the cases and controls are shown in No deviation of allele frequencies from Hardy–Weinberg equilibrium was found in both the cases and controls. As shown in Women with preeclampsia were further divided into four subgroups: early-onset mild preeclampsia (28 cases), early-onset severe preeclampsia (183 cases), late-onset mild preeclampsia (121 cases) and late-onset severe preeclampsia (122cases). The MAF was significantly different between early-onset severe preeclampsia and controls (P = 0.003). No significant differences were observed between the controls and other subgroups .Genotype frequency was further analyzed under additive, recessive and dominant models . SignifiIn the present study, a significant association between preeclampsia and rs11646213 was identified in a cohort of Han Chinese women. When the cases were divided into subgroups, there was a significant association between early-onset severe preeclampsia and the controls.The participants were all Han Chinese women in our study, so that there was no variation in genetic background. The T allele, which confers an increased risk of developing preeclampsia, was the minor allele. As shown in the GWAS blood pressure study, the A allele was identified as the minor allele in a European population CDH13, located on chromosome 16q24, encodes cadherin-13 and is highly expressed in the cardiovascular system CDH13CDH13 may be involved in the pathogenesis of preeclampsia through interactions with adiponectin.Preeclampsia is currently recognized as a syndrome rather than a definite disease. In this study, significant differences in genotype distribution were observed between early-onset severe preeclampsia and the controls after the cases were divided into subgroups. These results are in agreement with the theory that defective placental angiogenesis plays an important role in the development of early-onset preeclampsia, while late-onset preeclampsia generally has a normal placenta morphology CDH13 and preeclampsia need to be confirmed in a larger sample size composed of different ethnic groups. Second, the functional significance of the SNP rs11646213 remains unknown and molecular mechanisms regarding CDH13 in the pathophysiology of preeclampsia should be examined in future studies. Third, while we limited our study to only one genetic variant of CDH13, future studies should evaluate associations between preeclampsia and other genetic variants, particularly those that lack strong linkage disequilibrium with rs11646213.Several limitations should be considered. First, genetic factors that influence the pathogenesis of preeclampsia differ by ethnic group, and the association between In conclusion, we identified a significant association between rs11646213 and preeclampsia in Han Chinese women.Table S1Allele frequencies of rs11646213 in control, early-onset mild preeclampsia, early-onset severe preeclampsia, late-onset mild preeclampsia and late-onset severe preeclampsia subjects. PE: preeclampsia; MAF: minor allele frequency; OR: odds radio; 95% CI: 95% confidence interval.(DOCX)Click here for additional data file.


Twenty common genetic variants have been associated with risk of developing colorectal cancer (CRC) in genome wide association studies to date. Since large differences between populations exist, generalisability of findings to any specific population needs to be confirmed.The aim of this study was to perform an association study between risk variants: rs10795668, rs16892766, rs3802842 and rs4939827 and CRC risk in Croatian population.An association study was performed on 320 colorectal cancer cases and 594 controls recruited in Croatia. We genotyped four variants previously associated with CRC: rs10795668, rs16892766, rs3802842 and rs4939827.SMAD7 variant rs4939827 (18q21.1) was significantly associated with CRC risk in Croatian population. C allele was associated with a decreased risk, odds ratio (OR): 0.70 . Compared to TT homozygotes, risk was reduced by 34% in heterozygotes  and by 52% in CC homozygotes .Our results show association of rs4939827 with colorectal cancer risk in Croatian population. The higher strength of the association in comparison to other studies suggests population-specific environmental or genetic factors may be modifying the association. More studies are needed to further describe role of rs4939827 in CRC. Likely reason for failure of replication for other 3 loci is inadequate study power. It is t100,000) . Screeni100,000) .It has been estimated that over 30% of the variation in colorectal cancer susceptibility is attributable to genetic factors and majority of this seems to be due to multiple low risk mutations . To dateIt is known that the contribution of risk alleles to CRC risk may vary between populations, for example due to allele frequencies or specific linkage disequilibrium (LD) structure, or because of particular genetic and environmental backgrounds may modify the effect of the variants ,17. UndeThe role of risk loci in colorectal cancer occurrence is largely unexplored in Croatian population. In this paper, we set out to investigate the role of four risk loci: rs4939827, rs10795668, rs3802842 and rs16892766, on colorectal cancer risk in Croatian population.In the period from November 2008 to July 2009, a total of 338 colorectal cancer cases were recruited at departments of surgery in two hospitals in Zagreb, Croatia. Although both are located in Zagreb, these tertiary care hospitals attract patients from across the country. Data on age at diagnosis, sex, tumour histology and TNM stage were collected from CRC patients’ medical records and histopathology reports. Eighteen cases were excluded from the analysis because diagnosis of adenocarcinoma was not confirmed in histopathology report. Metastasis identified intraoperatively or confirmed by imaging methods were considered in evaluating disease progression. Colorectal cancer cases were classified into stages according to American Joint Committee on Cancer (AJCC) 7th edition .594 controls were selected from the ‘10,001 Dalmatians’ study, an ongoing genetic study in Croatia. This study includes approximately three thousand individuals from the general population of Split and from Vis and Korčula Islands in Croatia. Controls were picked randomly, but with constrains regarding age (+/- 5 years) and sex. Participants with personal history of malignant disease were excluded from the control group.Informed consent was obtained in writing from all participants. The study was approved by the appropriate ethics committees: the Ethics Committee of the University of Split Medical School and Ethical Board of the University Hospital Center Sestre milosrdnice.At recruitment, peripheral blood sample was collected from each subject and stored in EDTA coated vials. DNA was extracted using standard procedures. In cases, variants rs4939827, rs3802842, rs10795668 and rs16892766 were genotyped using the TaqMan SNP Genotyping Assay. DNA samples of controls were genotyped according to the manufacturer’s instructions on Illumina Infinium SNP bead microarrays (HumanHap300v1 or HumanCNV370-Quad). Genotypes were determined using Illumina BeadStudio software. Samples with a call rate below 97% were excluded. Variants rs4939827 and rs3802842 and were represented on the array and directly genotyped while rs10795668 and rs16892766 were imputed. Imputation with reference to HapMap CEU build 36 release 22 was performed using the software MACH v1.0.15 after exclusion of SNP with MAF <0.01, call rate <98% and HWE deviation p <10-6. Imputation quality was 99% for rs10795668 and 98% for rs16892766.2 test was used to assess deviations from Hardy-Weinberg equilibrium. Age- and sex-adjusted logistic regression analysis of CRC on allele dosage  was performed to assess the association between four variants and colorectal cancer risk, under the assumption of additive genetic model. Multiple testing corrected P-value of P<0.0125 was considered representative of statistical significance. R software was used for analyses and R package GenABEL was used for manipulating genetic data. χA total of 320  colorectal cancer cases and 594  controls were included in this study. Mean age of cases was 68.91 ± 10.08 y and of controls 66.12 ± 10.23 y. Where tumour site was known, 56% of cases had colon cancer and 44% had rectal or rectosygmoid tumour. Majority of cases had disease in AJCC stages 2 and 3 . The characteristics of cases and controls are detailed in All four variants conformed to the Hardy-Weinberg equilibrium. In the entire sample (cases and controls) allele frequency of T allele was 49.55%. In the logistic regression model we found statistically significant association between rs4939827 and CRC risk. The proportion of each of the rs4939827 genotypes CC, CT and TT was 29%, 49% and 22% in controls, respectively, and 21%, 47% and 32% in cases, respectively. C allele was associated with a decreased risk, odds ratio (OR) was 0.70 . Compared to TT homozygotes, risk was significantly reduced by 34% in heterozygotes  and by 52% in CC homozygotes  2.We next investigated the association between genotypes at rs4939827, rs10795668, rs3802842 and rs16892766 and tumour characteristics in an age and sex adjusted multivariate linear regression model. No statistically significant association was observed between selected variants and the following tumour characteristics: AJCC stage, tumour grade, tumour size or proportion of positive lymph nodes.In this study we found a significant association between SMAD7 variant rs4939827 (18q21.1) and risk of colorectal cancer in Croatian population. Compared to CC homozygotes, risk was increased by 58% in heterozygotes (CT) and by 111% in TT homozygotes.The mechanism by which rs4939827 may influence colorectal cancer involves inhibitory role of the SMAD7 protein in the transforming growth factor beta (TGF-β) signalling pathway ,20. TGF-per T=1.43). In a combined meta-analysis [In a comprehensive meta-analysis, we have previously shown a very high credibility of the association between rs4939827 and colorectal cancer . Howeveranalysis , found aanalysis  found anT=49.6%) were very similar to that of the other European populations , while prevalence of T allele is less frequent in Asian and African populations  (http://www.ensembl.org).Studies where higher OR was reported include  who founMoreover, replication studies can help identify population-specific environmental factors that modify the effect of risk loci. The relatively strong association between SMAD7 variant and CRC reported here, if verified, may be due to the particular genetic and/or environmental background specific for Croatian population. This finding warrants larger studies to explore the association further, as understanding the origin of the stronger effect in this population may help uncover mechanisms of colorectal cancer occurrence and progression, and may subsequently lead to translation of findings into more accurate risk prediction and better informed prevention should the interaction with environment be found.We failed to replicate associations between CRC risk and rs10795668, rs16892766 and rs3802842; however, this is may be due to the relatively small sample size -29 SimilLimitations of the study include relatively small sample size that may have contributed to our limited power to detect weak associations. Secondly, while controls were mainly from Split and islands of Vis and Korčula, cases were sampled from tertiary hospitals in Zagreb. However, this is not a major issue for this study: firstly, Croatian population is small and homogeneous; secondly, Zagreb and Split represent urban centres that attract immigration from other regions in Croatia, and lastly, patients would seek treatment in Zagreb due to perceived superiority of health care irrespective of their residence.In conclusion, a common variant rs4939827 is associated with colorectal cancer risk in Croatian population. Understanding differential effects in different populations may aid in uncovering disease mechanism and is important for the translation of results to risk prediction in different populations.


Previous studies have documented the increased cardiovascular risk associated with the use of some nonsteroidal anti-inflammatory drugs (NSAIDs). Despite this, many old NSAIDs are still prescribed worldwide. Most of the studies to date have been focused on specific oral drugs or limited by the number of cases examined. We studied the risk of new acute myocardial infarction (AMI) hospitalization with current use of a variety of oral and parenteral NSAIDs in a nationwide population, and compared our results with existing evidence.We conducted a case-crossover study using the Taiwan's National Health Insurance claim database, identifying patients with new AMI hospitalized in 2006. The 1-30 days and 91-120 days prior to the admission were defined as case and matched control period for each patient, respectively. Uses of NSAIDs during the respective periods were compared using conditional logistic regression and adjusted for use of co-medications.8354 new AMI hospitalization patients fulfilled the study criteria. 14 oral and 3 parenteral NSAIDs were selected based on drug utilization profile among 13.7 million NSAID users. The adjusted odds ratio, aOR , for risk of AMI and use of oral and parenteral non-selective NSAIDs were 1.42  and 3.35 , respectively, and significantly greater for parenteral than oral drugs (p for interaction < 0.01). Ketorolac was associated with the highest AMI risk among both of oral and parenteral NSAIDs studied, the aORs were 2.02  and 4.27  respectively. Use of oral flurbiprofen, ibuprofen, sulindac, diclofenac, and parenteral ketoprofen were also significantly associated with increased AMI risk. The results of the present study were consistent with the majority of evidence from previous studies.The collective evidence revealed the tendency of increased AMI risk with current use of some NSAIDs. A higher AMI risk associated with use of parenteral NSAIDs was observed in the present study. Ketorolac had the highest associated risk in both oral and parenteral NSAIDs studied. Though further investigation to confirm the association is warranted, prescribing physicians and the general public should be cautious about the potential risk of AMI when using NSAIDs. Non-steroidal anti-inflammatory drugs (NSAIDs) are commonly used medications for reducing inflammation and relief pain. Due to large population exposed to NSAIDs, the risk of serious adverse cardiovascular effect for patient taking NSAIDs is an area of concern, from both a clinical and public health perspective -9. Despith Revision, Clinical Modification, ICD-9-CM code), prescriptions, procedure, and examination ordered for every patient could be identified and traced by civil identification number. To comply with the personal electronic data-privacy regulation, personal identities were encrypted and all data were analysed anonymously. The study protocol was approved by the Research Ethics Committee of National Taiwan University Hospital.We used the claims database of Taiwan's National Health Insurance (NHI) for the present study. The NHI is a universal compulsory program launched in March 1995 by the Taiwan government. More than 98% of the total 23 million populations was covered by NHI at the end of year 2005 . Out-patIn order to compare the results of the present study with previous studies, we summarized previously published observational studies and reviews of randomized controlled trials (RCTs). The collection of observational studies was based on previous reviews ,11,14-16We used a case-crossover design to study the association between current use of NSAIDs and AMI hospitalization risk. This design is similar to a matched case-control study -51. One The target outcome of this study was new AMI hospitalization. We searched all the medical claims of NHI beneficiaries aged between 20 to 100 years old in order to identify the first hospitalized AMI (ICD-9-CM code 410) in 2006. To be eligible, patients needed to be continuously covered by NHI in 2005 and 2006. The date of the first AMI hospitalization was defined as the index date. Patients with any clinical visit, emergency-room visit or hospitalization for myocardial infarction (ICD-9-CM code 410 or 412) either 365 days preceding the index date or in 2005 were excluded. To increase the comparability of general health condition between the case and control period of a patient, those who had been hospitalized for any cause during 120 days prior to their index date were also excluded.The primary exposure of the present study included the COX-2 selective inhibitor and non-selective NSAIDs, which were reimbursed by NHI in Taiwan in 2005 and 2006. All prescriptions were searched for oral and parenteral single-active-ingredient NSAIDs and were classified according to the Anatomic Therapeutic Chemical (ATC) classification system . Drugs iThe association between risk of AMI hospitalization and use of NSAID was analysed by comparing the NSAID exposure status between the paired case and the control period of each patient. If a patient had not used an NSAID in the control period but had used it in the case period, there was a positive association between AMI hospitalization and current use of the NSAID. The association between AMI and NSAID would be negative if an NSAID was used in the control period but not in the case period. When a patient used an NSAID in both of case and control periods, or did not use an NSAID in either period, the association between AMI and the NSAID were neutral. Conditional logistic regression was used to analyse the paired data set with case and control period as dependent variables, and use or non-use of an NSAID as independent variables. Concomitant medications were considered as time-varying cofactors and were included to allow adjustment for potential within-patient time-varying confounding of effects. In addition to the primary analysis, we performed sensitive analyses by altering the case period to 8-30 days and the control period to 98-120 days prior to the index date; and by changing the control period to 61-90 days prior to the index date. We used the PHREG procedure, which is a feature of SAS statistics software , to calculate the crude and adjusted odds ratio (aOR) and their associated 95% confidence intervals (95%CI). A 2-sided p value of less than 0.05 was considered statistically significant.st January and 31st December 2006. We excluded 2,817 patients with previous MI diagnosis in 2005; 2,972 patients had been hospitalized for any cause during the 120 days prior to their index date; and the other 585 patients had outpatient visits with AMI diagnosis 365 days preceding their index date. The final analysis included 8,354 patients with a new AMI hospitalization , we identified 14,728 patients hospitalized with the principle diagnosis of AMI between 1The NHI drug-reimbursement list in 2006 shows 1009 oral and 119 parenteral single-active-ingredient NSAID items in ATC class M01A, "anti-inflammatory and anti-rheumatic products, non-steroids", excluding glucosamine (M01AX05). 13,699,038 patients received at least one NSAID prescription. Fourteen NSAIDs were selected for the present study according to the top 95 percentile user number. They were diclofenac , mefenamic acid , ibuprofen , naproxen , acemetacin , piroxicam , flurbirofen , sulindac , indomethacin , meloxicam , tiaprofenic acid , ketorolac , ketoprofen , and celecoxib . Celecoxib was the only COX-2 selective NSAID reimbursed during 2005 and 2006 in Taiwan. Though the number of patients using celecoxib was not large, the quantity used in terms of number of DDDs accounted for 6.15% of total NSAIDs DDDs used in 2006.Table Among the individual NSAIDs, ketorolac posed the highest risk for AMI in both of oral and parenteral NSAIDs. The aORs (95%CI) were 2.02  and 4.27  respectively. In addition to oral and parenteral ketorolac, the aORs for oral flurbiprofen, ibuprofen, sulindac, diclofenac, and parenteral ketoprofen were also statistically significantly greater than 1, ranging from 1.29 to 1.71 for oral drugs, and 2.34 for parenteral ketoprofen. The aORs for the rest of the other eight oral ns-NSAIDs and parenteral diclofenac were not statistically significant. No significantly protective effect to prevent new AMI hospitalization was observed for any NSAID studied.Table For individual NSAID in the subgroups, the aOR (95%CI) of celecoxib was significant only in patients with hypertension, which was 1.81 . The aORs of diclofenac were all significant, except in the low-dose-aspirin user subgroup. No aORs of individual oral NSAIDs were significant in the low-dose-aspirin user subgroup; but parenteral ketorolac had significant aOR of 7.47  for AMI in this subgroup. Most of the effects of NSAIDs on AMI were not substantially modified by hypertension diagnosis or use of low-dose-aspirin. Their 95%CIs in separate patient groups overlapped and the p values for testing interaction were greater than 0.15. The exceptions were observed for oral ns-NSAIDs overall and oral ketorolac in particular. Their effects on risk of AMI were higher in hypertensive patients than in patients without hypertension, and the p values test for interaction were 0.05 and 0.03 respectively. We did not find significantly modified effects by gender or age.Table The sensitivity analyses generated by altering case and control periods did not change the significant of the aORs of oral and parenteral ns-NSAIDs overall, and the higher risk with parenteral NSAIDs remained table . The assTable We found that the risk of new AMI hospitalization was increased with the current use of oral ns-NSAIDs overall. The risk was higher with parenteral ns-NSAIDs overall. The associations between AMI risk and individual oral ns-NSAID were mild to moderate, and not significantly increased for celecoxib with adjusted analysis. We did not find any specific NSAID with significantly protective effect to prevent new AMI hospitalization.The new AMI hospitalization risk in users of parenteral ns-NSAIDs overall was significantly higher than that observed in oral ns-NSAIDs users. This higher risk of parenteral ns-NSAIDs remained for subgroups categorized by status of hypertension diagnosis and use of low-dose-aspirin, and was consistent across sensitivity analyses.We found the use of ketorolac, in both oral and parenteral forms, was associated with the highest risk for new AMI hospitalization among all the NSAIDs. In 2006, 766,948 patients  received parenteral ketorolac, and more than one million DDDs (1 DDD = 30 mg) of injectable ketorolac were reimbursed by NHI  or recent (< 3 months) use of ns-NSAIDs or NSAIDs overall ,43,44,47 days or Among the individual ns-NSAIDs, for current us of naproxen: three previous studies reported significant reduction of AMI risk ,22,41; tExcept for naproxen, no significant reduction of AMI risk has been reported for current or recent use of other individual ns-NSAID in previous studies. This was also the case in our study. We did not observe a significant protective effect of any individual NSAID in preventing AMI hospitalization in any subgroup of patients. Nevertheless, significantly increased AMI risk with use of diclofenac has been repeatedly reported in previous studies ,41,43,47Consistent with four previous studies which found a significantly increased AMI risk with current use of ibuprofen ,25,37,47Flurbiprofen was another oral ns-NSAID associated with significantly elevated AMI risk in the present study. Few studies had previously evaluated this drug specifically. One study reported a non-significant 2.26 times increase of first-time AMI risk with flurbiprofen in a population based case-control study, but with only 22 users of this drug .One more oral ns-NSAID significantly associated with AMI risk in the present study was sulindac, with aOR (95%CI) of 1.44 . In the case-control study by Mangoni, users of sulindac and oxicams were pooled together and no association with AMI was found . HoweverWe did not find significant interaction effects between the use of NSAIDs and low-dose aspirin on AMI risk. Both significant and non-significant interaction results had been reported in previous studies ,27,31,48The interaction between NSAIDs use and hypertension in patients was also a concern . A fairlThe database used in this study covered the entire population of Taiwan. This not only provided large patient pool but also enabled follow up of each patient, thus minimizing the problem of case ascertainment. Owing to the large population pool, subjects recruited within a year were sufficient for most of the drugs studied, and more individual drugs could be analysed in one study. By limiting the recruitment period to one year, we were able to limit the potential influence of environmental changes, such as introduction of new drugs, change in medical practice, and epidemic trends of AMI in the community. The NHI reimburses a comprehensive list of prescription drugs, thus reducing the possibility and proportion of patients taking over-the-counter NSAIDs.An important issue for most non-randomized studies using cohort or nested case-control design to analyse AMI risk with use of NSAIDs is the challenge of selection of appropriate comparison subjects. The unmeasured risk factors of outcomes may differ between subjects in the comparison groups, leading to a confounded result. The case-crossover design of the present study utilized a within-subject comparison, removing the potentially confounding by between-subjects difference , has alsThe collective evidence revealed the tendency of increased AMI risk with current use of some NSAIDs. A higher AMI risk associated with use of parenteral NSAIDs was observed in the present study. Ketorolac had the highest associated risk in both oral and parenteral NSAIDs studied. Though further investigation to confirm the association is warranted, prescribing physicians and the general public should be cautious about the potential risk of AMI when using NSAIDs.The authors declare that they have no competing interests.All authors read and approved the final manuscript. The contributions of each author are:WYS conceived and designed the study, analysed and interpreted the data and drafted the manuscript; HCC had full access to all the data, took responsibility for the integrity of the data, carried out the statistical analysis and provided administrative support; STC had full access to all the data, took responsibility for the integrity of the data, carried out the statistical analysis and provided administrative support; CHC conceived and designed the study, analysed and interpreted the data, critically review and revised the manuscript; HWC conceived and designed the study, analysed and interpreted the data, provided administrative support, critically review and revised the manuscript; CWK conceived and designed the study, analysed and interpreted the data, critically review and revised the manuscript; MSL obtained funding, conceived and designed the study, analysed and interpreted the data, had full access to all the data, took responsibility for the integrity of the data, critically review and revised the manuscript and supervised the study.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2261/12/4/prepubsupplementary tables. There are 4 additional tables (table S1 to table S4) in the file to present the utilization pattern of the NSAIDs studied. Table S5 is the STROBE checklist for the present study.Click here for file


IRGM) gene may be associated with inflammatory bowel disease (IBD) by affecting autophagy. However, the genetic association studies on three common variants in IRGM gene  have shown inconsistent results.Polymorphisms in immunity-related GTPase family M (IRGM polymorphisms and IBD risk. Data were extracted and the odd ratios (ORs) and 95% confidence intervals (95% CIs) were calculated. Finally, we performed a meta-analysis of 25 eligible studies in 3 SNPs located at IRGM gene by using a total of 20590 IBD cases and 27670 controls. The analysis showed modest significant association for the rs13361189, rs4958847 and rs10065172 variants in Crohn’s disease (CD): the risk estimates for the allele contrast were OR=1.306 (1.200-1.420), p=5.2×10-10, OR=1.182 (1.082-1.290), p=0.0002, and OR=1.248 (1.057-1.473), p=0.009 respectively . When stratified by ethnicity, significantly increased CD risk was observed in Europeans, but not in Asians. Conversely, there was no association of rs13361189 or rs4958847 variant with risk of ulcerative colitis (UC).The PubMed and Embase were searched up to June 5, 2013 for studies on the association between three IRGM polymorphisms appear to confer susceptibility to CD but not UC, especially in Europeans. Our data may provide further understanding of the role of autophagy in the pathogenesis of CD.These results indicated that autophagy gene- Inflammatory bowel disease (IBD), a chronic inflammatory disease of the gastrointestinal tract, is usually classified into two clinical forms: Crohn’s disease (CD) and ulcerative colitis (UC) ,2. CD geATG16L1 and IRGM) may play an important role in the pathogenesis of IBD[IRGM) gene, located on chromosome 5q33.1, encodes a GTP-binding protein that induces autophagy, which is involved in elimination of intracellular pathogens[IRGM gene  with IBD has been recently reported [IRGM polymorphisms and CD susceptibility [Recent progress in the genetics of IBD has advanced understanding of disease pathogenesis. GWAS meta-analysis identified 71, 47 and 163 susceptibility loci of CD, UC and IBD, respectively. These genes involved in intestinal barrier function (GNA12 and LAMB1), transcriptional regulation (NKX2-3 and IRF5) and immune response (IL23R and IL12B). Recently, studies in animal models and IBD patients suggested that autophagy related genes  susceptibility. Meta-analysis is a proper method to deal with these ambiguities and overcome the problem of small sample sizes and inadequate statistical power in different genetic studies. In the present study, we performed a meta-analysis of all eligible case control and cohort studies to clarify the associations between three common polymorphisms  in the IRGM’, ‘polymorphism’ or ‘variant’, ‘rs13361189’, ‘rs4958847’ or ‘rs10065172’, and ‘single nucleotide polymorphism’ or ‘SNP’( the last search update was 5 June 2013). In addition, the reference lists of all retrieved articles were checked for additional potential studies. A study was included in the analysis if (1) reported the relationship between the polymorphisms of IRGM rs13361189, rs4958847 or rs10065172and the risk of IBD; (2) the genetic information of included studies was from unrelated populations (studies of which the design was not based on family data). Major reasons for exclusion of studies: (1) no control population; (2) studies that contained overlapping data; (3) comments, letters, review articles, or articles only with an abstract. Additionally, when a study reported the results on different subpopulations or panels, we treated them as separate studies in the meta-analysis.Electronic searches in Medline, Embase, CNKI  and Chinese Biomedicine databases were performed using the following search terms: ‘Inflammatory Bowel Disease’ or ‘IBD’, ‘Crohn’s disease’ or ‘CD’, ‘ulcerative colitis’ or ‘UC’, ‘The following data was extracted independently from each study by two authors: first author, journal, year of publication, country of origin, ethnicity of the individuals involved , genotype frequency, sex and mean age in cases and controls. Of the studies with the overlapping data of the same population resource, we selected the most recent ones with the largest number of participants. If the article did not provide sufficient genotype distribution, the corresponding author was contacted for the detailed data. In addition, disagreements were resolved by discussion between the two investigators.IRGM polymorphisms and CD or UC risk was evaluated by the odds ratios (ORs) with 95% confidence intervals (CIs). For the rs13361189 polymorphism, we examined the allelic effect of C  versus T , and also examined the contrast of CC versus TT, CT versus TT, CC+CT versus TT (dominant model), as well as CC versus CT+TT (recessive model). Similar models were analyzed for the rs4958847 and rs10065172 polymorphisms. The association between rs10065172 and UC risk was not evaluated for the lack of sufficient data. The significance of the pooled OR was determined by the Z-test; and the P values were adjusted using Bonferroni correction by the number of compared SNPs. (P=0.05/3=0.017) In addition, for each genetic contrast, stratified analysis was performed according to ethnicity. The Hardy-Weinberg equilibrium (HWE) in the control group was assessed, and P<0.05 was considered as significant disequilibrium.The strength of the association between 2 test. Heterogeneity was considered significant when P<0.10, and then the random effect model was applied for meta-analysis, otherwise, a fixed-effects model was used [2 takes values between 0% and 100% with higher values denoting a greater degree of heterogeneity [The heterogeneity between studies was assessed by Chi-square based Q test  and I2 twas used . I2 takeogeneity . In addiCumulative meta-analyses were carried out for all three variants in association with CD and two variants (rs13361189 and rs4958847) with UC to evaluate the trend of the genetic risk effect (OR) of the allele contrast as evidence accumulating over time. To assess the stability of the results, sensitivity analysis was carried out after sequential removal of each study or by excluding those studies deviated from HWE.Publication bias was investigated using graphical evaluation of funnel plots. However, the funnel plot may be not considered strictly as a test of publication bias. Then, the Egger’s test was used to provide statistical evidence of funnel plot symmetry . If signIRGM polymorphisms  and IBD (UC or CD) were included in this meta-analysis. The literature review identified 62 articles in PubMed, Embase, CNKI and Chinese Biomedicine databases that met the search criteria. The abstracts and full articles of the retrieved studies were read to assess their appropriateness for meta-analysis. Finally, a total of 23 relevant articles with IRGM polymorphisms was extracted from 25 studies, 4 studies [A list of details of the studies included in the meta-analysis was provided in  studies ,28,30,31 studies  was incl studies ,21, the -13) of rs13361189 C allele. Similar result was observed in rs4958847 (p=1.8×10-10). For rs10065172 variant, as only one study carried out in Asians was included, one sample T-test was used to compare the differences of allele frequencies between Asian and European controls (p=9.9×10-7).  for rs13361189 variant, 34397 individuals (10924 cases and 23473 controls) for rs4958847 variant, and 16972 individuals (6766 cases and 10206 controls) for rs10065172 variant. We observed a wide spectrum of the rs13361189 C allele and rs4958847 A allele frequencies across different ethnicities. Compared with Europeans controls , Asian controls carried a higher frequency . -10, OR=1.182 (1.082-1.290), p=0.0002 and OR=1.248 (1.057-1.473), p=0.009 respectively, significant even after Bonferroni correction). Moreover, significant correlation was also found in the three polymorphisms under other genetic models . (models). -25, significant after Bonferroni correction) However, these associations were not observed in the Asian populations (CC+CT vs TT: OR=1.199 (0.987-1.455), p=0.068). Similarly, statistical association was observed between rs4958847 or rs10065172 variant and CD risk among European population. (AA+AG vs GG: OR=1.269 (1.147-1.403), p=3.6×10-6, still significant after Bonferroni correction) and TT+TC vs TT: OR=1.298 (1.008-1.673), p=0.043, non-significant after Bonferroni correction) In addition, the OR for rs13361189 T>C was 1.565 (1.218-2.010) in carriers of two risk C alleles compared with non-risk allele carriers (CC vs TT), which was higher than the risk of one T allele carriers ), suggesting a dose–response with increasing number of the variant allele. The same pattern was seen for either rs4958847 or rs10065172 variant. In the subgroup analysis by ethnicity, for rs13361189 T>C, significantly increased CD risk was found among European populations in the allelic and all genetic models. (CC+CT vs TT: OR=1.421 (1.329-1.519), p=5.0×10IRGM gene and the risk of UC were shown in  Meta-analysis findings of associations between rs13361189 and rs4958847 in the IRGM SNPs in the total analysis of CD. , rs4958847 variant , or rs10065172 variant (P=0.479 for TT+TC vs CC in CD). Toxoplasma gondii and Listeria monocytogenes [-8, meeting genome-wide significance). Parkes et al [IRGM variants (rs13361189 and rs4958847) associated with CD in two different British cohorts. Thus, IRGM may appear to be a good candidate for IBD. However, the results of associations between three IRGM variants  and risk of IBD were contradictory. Therefore, we saw the need to perform pooled analyses with larger sample size by summarizing previous case-control studies in order to understand the association between IRGM variants and IBD risk better.IRGM gene, located on chromosome 5q33.1, plays an important role in autophagy. Autophagy has a central function in physiological and pathological processes, involving in innate and adaptive immunity by delivering intracellular pathogens and other antigens. Singh et al demonstrated that IRGM can induce autophagy to eliminate intracellular mycobacterium tuberculosis . Moreoveytogenes . The preytogenes . In addiytogenes . In the es et al  found tw2>0.8). The results showed that rs13361189 was in perfect LD with 2 other IRGM SNPs  in Europeans. However, rs4958847 was not in LD with 3 other IRGM SNPs . Our meta-analysis showed significant susceptibility of CD from rs13361189 in the overall population in all genetic contrasts. When stratified by ethnicity, a significant association with rs13361189 was observed in European population, but not in Asians. Similar results were also found between the rs4958847 or rs10065172 variant and risk of CD in overall and European population. It is widely accepted that genetic markers in predisposition to IBD vary across ethnic groups. For instance, nucleotide oligomerization domain 2 (NOD2) polymorphisms have been strongly association with CD in Europeans ,46, but NOD2 and ATG16L1 were associated with CD, but not with UC [Overall, no significant association between rs13361189 or rs4958847variant and susceptibility to UC was found in this meta-analysis in any genetic model. To date, there was lack of association of these two SNPs with UC in all the individual studies. Recently, twin studies and familial clustering of cases suggested that genetic factors were likely to play a more prominent role in CD than in UC [ with UC ,51. 2. Moreover, the heterogeneity was remarkably decreased among Asian and European population, , which may be attributed that IBD is a complex disease and different genetic backgrounds or different environments existed among different ethnicities. Moreover, the sample size could explain 63.28% of τ2 under dominant model. In addition, the pooled OR did not change in the sensitivity analysis by excluding studies departed from HWE.Heterogeneity was significant for the most comparisons of rs13361189 polymorphism in overall population. To identify the source of heterogeneity, meta-regression and subgroup analysis were carried out. We found that ethnicity was identified as a potential source of between-study heterogeneity. Meta-regression indicated that ethnicity could explain 41.37% of τOur meta-analysis significantly increased statistical power by pooling data from different studies, while several limitations should be considered in the present meta-analysis. First, only 2 and 3 studies were performed in Asians for rs4955847 and rs13361189 variants, respectively. Therefore validation of association is required in other population. Second, significant heterogeneity between studies was detected in the current meta-analysis, whereas difference in ethnicity was identified as potential sources of heterogeneity. Third, gene–environment and gene-gene interactions were not analyzed because of insufficient data. IRGM polymorphisms in CD or UC. To identify the exact role of IRGM polymorphisms in the pathogenesis of CD, more studies such as animal disease modeling are of great importance.In conclusion, despite these limitations, our meta-analysis still yields statistically significant results. The present data synthesis indicated that rs13361189, rs4958847 and rs10065172 were considered to be risk factors of CD in Europeans but not of UC. In addition, subgroups analysis suggested that this increased risk may be ethno-specific. Further studies in other ethnic groups (e.g. Asians and Africans) are needed to clarify possible roles of Checklist S1PRISMA checklist.(DOC)Click here for additional data file.Figure S1OR estimates with the corresponding 95% CI for the associations between IRGM polymorphisms ((A) rs13361189 and (B) rs4958847) and the risk of UC (dominate model).(TIF)Click here for additional data file.Figure S2Sensitivity analysis on the associations between IRGM polymorphisms ((A) rs13361189 and (B) rs4958847) and UC risk (dominate model).(TIF)Click here for additional data file.Figure S3Cumulative meta-analysis: pooled OR with the corresponding 95% CI at the end of each year information step is shown for IRGM polymorphisms ((A) rs13361189 and (B) rs4958847) and risk of UC (dominate model).(TIF)Click here for additional data file.Figure S4Funnel plots of the association between IRGM polymorphisms ((A) rs13361189, (B) rs4958847, and (C) rs10065172) and CD risk (dominant model).(TIF)Click here for additional data file.Figure S5Funnel plots of the association between IRGM polymorphisms ((A) rs13361189 and (B) rs4958847) and UC risk (dominate model).(TIF)Click here for additional data file.


P = 0.04), annual number of hospitalizations >3 , average length of hospitalization >7 days , and a pathological transthoracic echocardiography (TTE) . Although the retrospective design and small sample size are weaknesses of the present study, these results are consistent with those of previous studies and allowed identifying associated factors such as a pathological TTE.A matched case-control study was performed in order to identify some associated factors for ACS or to confirm the published data. Controls were children hospitalized during the same period for pain crisis who did not develop an ACS during hospitalization. Between January 2006 and October 2010, there were 24 episodes of ACS distributed among 19 patients (8 girls and 11 boys). The median age was 7.5 years (range: 3 to 17 years) for the cases and 7 years (range: 3–18 years) for the controls. Four cases and 11 controls were treated with hydroxyurea (HU). In 75% of the cases, the ACS had arisen 24–72 hours following admission. The independent factors associated with ACS were average Hb rate <8 g/dL (OR = 4.96, 95% CI = 1.29–27.34, and  The acute chest syndrome (ACS) is a complication of SCD characterized by pleuritic chest pain, fever, rales on lung auscultation, and pulmonary infiltrates on chest X-ray [Sickle cell disease (SCD) is a major public health concern in French Guiana, a French region with 230,000 inhabitants located in South America . The incst X-ray . In 1979We report here the results of a case-control study of risk factors for ACS in children with SCD in French Guiana, in order to find some associated factors for ACS or to confirm the published data. We hypothesized that HbSS, age, high Hb level, and high steady-state leukocyte count could be risk factors for ACS. This matched case-control study concerned all cases of ACS hospitalized in the pediatric unit in French Guiana from 2006 to 2010. The cases were children hospitalized between January 2006 and October 2010 for pain crisis and who developed an ACS. The controls were children hospitalized during the same period for pain crisis and who did not develop an ACS during hospitalization. Each episode of ACS was matched on age, gender, and year of diagnosis.P value of 0.05 or less was considered as statistically significant. The data were entered into Microsoft Excel 2007 and analyzed using R.2.10.0  statistical software. All the factors numbered in The transthoracic echocardiography (TTE) was performed by a single pediatrician cardiologist, at baseline when the child was in a healthy state, during the annual evaluation. Patients with a pathological TTE were followed every six months by the same pediatrician cardiologist. All the TTE were obtained at true baseline and not during admissions. These TTE showed the following anatomical pathologies: an enlargement of the left heart chambers associated with an elevation in blood volume in seven cases and two controls and elevation in left ventricular myocardial indices in two cases and two controls. The Commission Nationale Informatique et Libertés approved our data collection. The factors associated with ACS were analyzed by logistic regression based on odds ratios (OR). For all tests performed, a Between January 2006 and October 2010, there were 24 episodes of ACS distributed among 19 patients (8 girls and 11 boys). One patient developed 3 events of ACS; 3 presented 2 events; and 15 patients presented 1 event of ACS. The median age was 7.5 years (range: 3 to 17 years) for the cases and 7 years (range: 3–18 years) for the controls (Mann-Whitney test). These two groups did not differ in age.β° thalassemia, and 1 HbSC. Among the controls, there were 45 HbSS, 12 HbSβ° thalassemia, 10 HbSβ+ thalassemia, and 9 HbSC. The more frequent hospitalization, longer hospital stays, lower Hb, higher % HbS, and earlier presentation in the cases may be explained, at least in part, by the difference in genotype and the severity of sickle cell anaemia.Four cases and 11 controls were treated with hydroxyurea (HU). The results are shown in In 75% of the cases, the ACS had arisen 24–72 hours following admission. All patients received rehydration, oxygen therapy, and pulmonary physiotherapy using stress spirometry and triple antibiotic therapy . In 16 cases, patients received a single red blood cell transfusion and in six other cases, the red blood cell transfusion was followed by a partial exchange transfusion . The target haemoglobin S value was under 30% and haemoglobin level 80–90 g/L, and in any of the cases, we obtained them. The more frequent hospitalization, longer hospital stays, lower Hb, higher % HbS, and earlier presentation in the cases may be explained, at least in part, by the difference in genotype and the severity of sickle cell anaemia. Two cases died. The cases who died did not receive a transfusion. The first was a 6-year-old girl in severe vasoocclusive crisis with fever and severe anemia (Hb of 4 g/dL), who presented 8 hours later with a frank ACS and whom endotracheal intubation was unsuccessful. The second was a 4-year-old girl also with frank ACS and bilateral pleural effusion, who died despite successful endotracheal intubation. The two cases who died had received a single dose of ceftriaxone. The proportion of death was high and due in part to low access to care in foreign patients. Although health is free of charge in French Guiana, the prevention is complicated by the fact that a number of persons are illiterate or do not understand the language. However, the health priorities of immigrants are often overridden by daily struggles to obtain food, shelter, and papers. Due to the low power of our study, certain factors such as lower Hb, higher % HbS, and earlier presentation in the cases as well as the transcranial Doppler, that were significant in bivariate analysis, were not statistically significant in multivariate analysis. However, the effect of including several episodes in single individuals was not analysed because of the small sample of our study.P = 0.003), average length of hospitalization >7 days , average Hb rate <8 g/dL , and a pathological TTE  were independent associated factors for ACS. The TTE was performed to detect any abnormalities such as left heart chambers abnormalities and intracardiac shunts, including foramen ovale. According to a few studies [Annual number of hospitalizations >3 (OR = 5.44, 95% CI = 3.59–8.21, and   studies , intraca