Backgrounds. Outcome of childhood malignancy has been improved mostly due to the advances in diagnostic techniques and treatment strategies. While methotrexate (MTX) related polymorphisms have been under investigation in childhood malignancies, many controversial results have been offered. Objectives. To evaluate associations of polymorphisms related MTX metabolisms and clinical course in childhood lymphoid malignancies. Method. Eighty-two acute lymphoblastic leukemia and 21 non-Hodgkin's lymphoma children were enrolled in this study. Four single nucleotide polymorphisms in 2 genes  and SLCO1B1 (rs4149056/c.521T>C/p.V174A and rs11045879/c.1865+4846T>C)) were genotyped by Taqman PCR method or direct sequencing. Clinical courses were reviewed retrospectively. Results. No patient who had the AC/CC genotype of rs1801131 (MTHFR) had relapsed or died, in which distribution was statistically different among the AA genotype of rs1801131 (P = 0.004). Polymorphisms of SLCO1B1 (rs11045879 and rs4149056) were not correlated with MTX concentrations, adverse events, or disease outcome. Conclusions. Polymorphisms of MTHFR (rs1801131) could be the plausive candidate for prognostic predictor in childhood lymphoid malignancies. Childhood cancer is a rare disease affecting 1 in 70,000 children aged 14 years and younger . LymphoiMTHFR polymorphisms, rs1801133  and rs1801131 , that have been shown to have lower enzyme activity when they carry mutant alleles [Methotrexate (MTX) is one of the key drugs for cancer treatment and a proven critical component for pediatric ALL and NHL , 4, 5. MMTHFR rs1801133 was associated with an increased risk of relapse events but not associated with MTX concentrations nor adverse events and that rs1801131 was not associated with altered risks of relapse nor toxicity in 520 pediatric ALL patients by the Children's Oncology Group [MTHFR rs1801133 resulted in a better overall survival rate in 126 Brazilian pediatric ALL patients treated with MTX [MTHFR c.T677A1298 haplotype) had a lower event free survival [MTHFR rs1801133 and C allele of MTHFR rs1801131 had higher relapse ration [A previous study reported that the T allele of with MTX , and othsurvival , and T ae ration .SLCO1B1 transfected cells were proven to uptake MTX in vitro, as well as other compounds such as estradiol, bilirubin, bile acids, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA), rifampicin, angiotensin-converting enzyme inhibitors, and the active metabolite of irinotecan, SN-38 [SLCO1B1 polymorphisms  altered in transport of substrates in vitro [SLCO1B1 polymorphisms rs4149056  transfected HEK cells, decreasing transporting activities [SLCO1B1 polymorphism rs11045879 (c.1865+4846T>C) and rs4149056 were shown to have lower MTX elimination in a genome-wide-association-study (GWAS), whose cohort mostly consisted of Caucasians and a limited number of Asians [SLCO1B1 were confirmed associations with a higher MTX concentration at 72 hrs among Spanish B-ALL children [SLCO1B1 polymorphisms ) and MTX concentration nor disease status [Transporters such as adenosine triphosphate-binding cassette (ABC) transporters and organic anion transporters were also reported to act for MTX disposition . Solute n, SN-38 –20. Tiroin vitro . Kameyamf Asians . These pchildren , 23. Jape status .MTHFR polymorphisms and MTX toxicity in pediatric ALL patients, results derived from these studies are conflicting. Study populations were mostly Caucasians and reports from Asians were limited [Although many studies have been conducted to investigate associations between  limited , 25. All limited , 26, 27.MTHFR (rs1801133 and rs1801131) and 2 more polymorphisms in SLCO1B1 (rs4149056 and rs11045879) in Japanese ALL/NHL patients treated with HD-MTX and examined the relationship between genotypes and prognosis/adverse events.In the present study, we genotyped 2 polymorphisms in MTHFR (rs1801133 and rs1801131) and SLCO1B1 (rs4149056 and rs11045879) and prognosis or clinical course including MTX concentrations in Japanese children who developed lymphoid malignancy treated with HD-MTX.To investigate associations of polymorphisms of Eighty-two acute lymphoblastic leukemia (ALL) and 21 non-Hodgkin lymphoma (NHL) children were enrolled in the present study. All patients were treated at two main and exclusively pediatric malignancy treating regional hospitals in Ibaraki prefecture  between November 1993 and November 2012. Patients' characteristics are shown in Informed consent was obtained from each parent/guardian or patient. The study was approved by the ethics committee of the University of Tsukuba in accordance with the Ethical Guidelines for Human Genome/Gene Analysis Research of the Ministry of Health, Labor and Welfare of Japan and the Declaration of Helsinki.2/12 hrs or 24 hrs, or 5 g/m2/24 hrs with folic acid rescue following the protocol of the Tokyo Children's Cancer Study Group (TCCSG) L99-15 [All patients received intravenous MTX continuously at 3 g/m) L99-15 , ALL-BFM) L99-15 , or NHL ) L99-15 .μmol/L.Monitoring of MTX concentration in plasma was carried out every day until the concentration was below 0.1 http://ctep.cancer.gov/). The highest grade of toxicity observed for each patient during the MTX therapy period was recorded. Data were collected including vomiting, diarrhea, serum hepatic enzyme (ALT), serum bilirubin and renal toxicity (serum creatinine), and MTX concentrations at 48 and 72 hrs after infusion. MTX levels were considered high if the concentration was above 1.0 μmol/L at 48 hrs or 0.1 μmol/L at 72 hrs. Above 1.5 mg/dL was considered as hyperbilirubinemia.Toxicity data were retrospectively collected objectively, blinded genotypes, from the patients' medical files. Toxicity was graded according to the Common Terminology Criteria for Adverse Event (CTCAE) v4.0 released from the Cancer Therapy Evaluation Program of the National Cancer Institute  were categorized as having global toxicity.DNA was extracted using Genomic DNA Isolation Kit  from 0.5 to 2 mL peripheral blood, bone marrow, or paraffin-embedded bone marrow/tissue in complete remission following the manufacturer's instructions.MTHFR  and SLCO1B1  were genotyped using the TaqMan Assay-on-Demand SNP Typing System  following the manufacturer's instructions. PCR was performed on a 384-well format with 5 ng of DNA each, and automatic allele calling was performed using ABI PRISM 7900HT data collection and analysis software, version 2.2.2 (Applied Biosystems).Polymorphisms of The accuracy of the genotyping for rs1801131 in children who had relapsed or died was confirmed by direct sequence using the primers 5′-TTTGGGGAGCTGAAGGACTA-3′ (forward) and 5′-CTTTGTGACCATTCCGGTTT-3′ (reverse) as reported by Shimasaki et al. [http://www.ncbi.nlm.nih.gov/) and Genome Medicine Database of Japan  which consisted of healthy Japanese individuals and have been genotyped using Illumina Sentrix HumanHap550 Genotyping BeadChip and InfiniumHD 610-Quad BeadChip , and principal component analysis has been performed to remove relatives, duplications, and Han Chinese . The genetic effects of the association between the case-control status and each individual SNP were assessed by chi-square test or Fisher's exact test.Deviation from Hardy-Weinberg expectations was examined by chi-square test. Control genotype frequencies were obtained from HapMap database  that each patient experienced during all HD-MTX courses and polymorphisms of To confirm the suitability of MTX concentrations as a toxicity marker, the associations between different toxicity markers  and MTX plasma concentration at 48 and 72 hrs among evaluable 149 courses conducted for ALL in total were examined by chi-square test or Fisher's exact test.P value <0.05 was considered statistically significant.Analysis was conducted using SPSS, version 12.0 . A Children enrolled in our study consisted of 82 patients diagnosed as ALL  and 21 children diagnosed as NHL . The number of patients who were in their first complete remission was 89, relapsed 8, and died 6. The average age at diagnosis was 7.4 (0.2–19.2) years old. There were 62 boys and 41 girls. Patients' characteristics are shown in MTHFR and SLCO1B1 in patients and controls Hapmap JPT obtained from the NCBI database and GeMDBJ. Deviation from Hardy-Weinberg expectations was examined by chi-square test and was applied to each of SNPs . Differences of allele frequencies were assessed by chi-square test and there were no differences with control data. Among Hapmap JPT, the P value for rs1801133 was 0.442, rs1801131 was 0.924, rs11045879 was 0.896, and rs4149056 was 0.066. Among GeMDBJ, the P value for rs1801133 was 0.885, rs1801131 was 0.765, and rs4149056 was 0.408.MTHFR c.1298AC/CC genotype), and no children who relapsed/died carried this genotype, in which frequency was significantly different (P = 0.004). MTHFR c.1298A>C (rs1801131).The outcome and each of polymorphisms are shown in 2 HD-MTX courses following the treatment protocol of TCCSG L99-15 for standard or high risk ALL patients totaled 149. For assessing the associations of adverse events during each MTX administration and each genotype, we analyzed these homogeneously treated 149 courses of weekly HD-MTX (MTHFR c.1298AA) tended to correlate to MTX concentration of higher than 1.0 μmol/L at 48 hrs after administration but not statistically significant (P = 0.06), and no other genotypes were correlated to MTX concentrations.All assessable weekly 3 g/my HD-MTX . CoursesMTHFR c.677CC) developed Common Terminology Criteria for Adverse Event (CTCAE) v4.0 more than grade 1 ALT elevation, whereas 41 of 100 courses undertaken for children with at least one T allele of rs1801133 (MTHFR c.677CT/TT) developed elevated serum liver enzyme (P = 0.006). Thirty-eight of 96 courses undertaken for children with the AA genotype of rs1801131 (MTHFR c.1298AA) resulted in elevated serum liver enzyme, 11 of 52 with at least one C allele of rs1801131 (MTHFR c.1298AC/CC genotype) resulted in ALT elevation (P = 0.036). Only 8 of 32 courses conducted with patients who carried the CC genotype of rs1801133 (MTHFR c.677CC) resulted in the next MTX administration being delayed more than 5 days due to adverse events such as serum hepatic enzyme, renal toxicity, or infection, as 33 of 69 courses for patients who carry at least one T allele of rs1801133 (MTHFR c.677CT/TT genotype) resulted in the next MTX administration being delayed which had been planned to have been conducted weekly (P = 0.030).Only 9 of 49 courses undertaken for children with the CC genotype of rs1801133 (SLCO1B1 (rs11045879 and rs4149056)  Tables  and 3.P < 0.001) and delayed weekly MTX administration (P = 0.013).MTX concentrations and adverse events were analyzed by chi-square test . ElevateHigh dose methotrexate is highly effective for broad cancers including lymphoid malignancies such as leukemia and lymphoma. However, its adverse events differ among each ethnicity and could significantly affect its clinical course. Previously, genetic polymorphisms of MTX pathway protein were reported as one of the predictable values.P = 0.004). Previous studies using Caucasian ALL patients reported that the T allele of rs1801133 was correlated with poor prognosis, but the polymorphism of rs1801131 was not associated with the prognosis of the patients [MTHFR c.T677A1298 haplotype) had a lower event free survival among Caucasians [With recent excellent advances in the prognosis among childhood lymphoid malignancy, the rate of patients who relapsed or died is very limited. It is vitally important to detect the factors that influence the prognosis in this limited number of poorer prognostic patient cohort other than factors that have already been thoroughly examined. In our study, all children who had relapsed or died carried the AA genotype of rs1801131  was reported as a candidate predictor of MTX lower elimination, but its results are controversial and many different results have been reported [SLCO1B1 (rs11045879 and rs4149056). Lopez-Lopez et al. confirmed this result in Spanish population [SLCO1B1 polymorphism in pediatric lymphoid malignancies had not been conducted previously.Previously, rs1801133  tended to have a higher MTX concentration at 48 hrs; however, this association did not reach statistical significance. Polymorphisms in SLCO1B1 (rs11045879 and rs4149056) were not correlated to MTX concentrations at 48 hrs . The discrepancies among studies previously conducted among Caucasians might be caused by differences of genetic backgrounds, lifestyles, or treatment protocol. This is retrospective study, so in the future a prospective research for Japanese children is warranted.MTX concentration at 48 hrs was one of the strong predictors to develop a delay in the next course of MTX administration and serum creatinine elevation . In our MTHFR (rs1801133 and rs1801131) were associated with serum ALT elevation . Patients who carried at least one T allele of rs1801133 (MTHFR c.677CT/TT genotype) were also associated with more than 5 days of delay of the next MTX administration, which was planned to undergo weekly (P = 0.03). Patients with adult acute lymphoblastic leukemia carrying at least one T allele of rs1801133 (MTHFR c.677CT/TT genotype) were associated with developing ALT elevation as CTCAE v.4.0 grade more than 1, which was the same result as reported previously [MTHFR c.1298 AC) showed lower adverse event [MTHFR c.1298AC) required higher MTX concentrations in order to inhibit 50% of the control TS activity ex vivo [Polymorphisms in eviously , 31. Chi ex vivo . These rSLCO1B1 and MTX eliminations were discovered by the GWAS study, which mostly consists of ancestry of European or African descent and a small number of Asian descent [P = 0.529 for rs11045879, P = 0.413 for rs4149056)  . The difMTHFR and rs11045879 and rs4149056 of SLCO1B1) and adverse events during HD-MTX treatment and prognosis in Japanese childhood lymphoid malignancies.We conducted an association study of polymorphisms (rs1801133 and rs1801131 of MTHFR c.1298AC/CC genotype) had not relapsed nor died. The CT/TT genotype of rs1801133 (MTHFR c.677CT/TT genotype) resulted in higher serum hepatic enzyme and delayed administrations during weekly high dose MTX.Patients that carried polymorphisms of rs1801131 (SLCO1B1 polymorphisms (rs11045879 and rs41490586) in Japanese children were assessed for the first time and found to have no correlations with any adverse events, MTX concentrations or outcome.Relations of


The relationship between congenital hypothyroidism (CH) and multiple pregnancy has not been fully studied in Japan.Complete nationwide data of assisted reproductive technology (ART) in Japan from 2005 to 2009 presented by the Japan Society of Obstetrics and Gynecology, which include information on CH and birth defects were used for analyses. Diseases were reclassified according to the International Classification of Diseases, tenth edition . The relative risk (RR) and 95% confidence interval (CI) of the incidence rate for CH was calculated with singletons as the reference group. Additional birth defects with CH were examined. The probandwise concordance rate of multiples and recurrence risk ratio were calculated.There were 18 patients with CH, consisting of 12 singletons and 6 multiples. The incidence rates of CH per 1,000 live births in singletons and multiples are 0.14  and 0.31 , respectively. The incidence rate was more than twofold higher in multiple births than in singleton births, but the difference was not statistically significant . Additional birth defects were present in three patients with CH (17%=3/18). The multiples were affected by other birth defects more frequently (33%=2/6) than the singletons (8%=1/12). The six multiple-births patients were derived from one concordant twin pair, which consisted of two twin patients; three discordant twin pairs, which consisted of three twin patients; and one discordant triplets set, which consisted of one triplet patient. Thus, the probandwise concordance rate was 33.3% (=2/(2+4)). The estimated recurrence risk ratio was 1976  or 609 .CH was more frequent in multiples compared to singletons. A higher percentage of additional birth defects was also observed in multiples compared to singletons. The familial aggregation of CH was suggested. Congenital hypothyroidism (CH) is defined as thyroid hormone deficiency present at birth and is classified into permanent and transient CH . An exacSeveral risk factors of CH have been presented in epidemiologic studies. Females are consistently reported to more frequently suffer from CH than males ,8. The aMany case studies showed that CH occurs sporadically , althougGiven these circumstances, assisted reproductive technology (ART) data present a unique opportunity for twin study . The perThe method for collecting data is described in detail elsewhere . Almost The author used case reports on birth defects or congenital anomaly data from 2005–2009 as the initial information for the present secondary data analyses. The mean response rate throughout the 5 years was 99.0% , meaning that almost a complete database reflecting the current situation of ART in Japan could be constructed.All live births after ART were analyzed in the present study. The types of defects were reclassified according to the International Classification of Diseases, tenth edition . Diseases that were classified in the category of ICD-10 code Q00-Q99  were regarded as birth defects in the present study. CH was classified as part of E00  or E03 (other hypothyroidism). The present data did not allow the author to distinguish between permanent CH and transient CH. Thus, all types of CH were treated as CH in the present study. The total number of ART live births, singletons and multiples, were available from 2007 to 2009. The author estimated the number of ART singletons and multiples between 2005 and 2006 using approximation formulae .All patients with CH were listed with their obstetric data and neonatal outcome to further examine the features of CH patients. The frequency of additional birth defects in neonates with CH was calculated. Next, the relative risk (RR: the incidence rate of multiples/singletons) with the corresponding 95% confidence interval (CI) was calculated with singletons as the reference group.Finally, the familial aggregation of CH was analyzed. The probandwise concordance rate  was calcA widely used measure of familial aggregation is the sibling recurrence risk ratio (RRR), defined as the ratio of the risk of disease manifestation, given that one’s sibling is affected, compared with the disease incidence rate in the general population ,32. The Statistical analysis was conducted using Microsoft Excel 2010 and SAS for Windows ver. 9.2.Demographic and perinatal outcome data of all neonates with CH are presented in Table The incidence rate of CH per 1,000 live births was 0.17 . The rate of CH was more than twofold higher in multiple births  than singleton births , but the difference was not statistically significant . The proportion of multiple births observed in the CH patients (33%=6/18) was twofold higher than that estimated in the total ART population  in the same period (2005–2009).The calculated concordance rate and RRR are presented in Table http://www.boshiaiikukai.jp/img/milk/kensajokyoH22.pdf (In Japanese). Accessed December 23, 2012). The lower incidence rate of CH in the present study might be attributed to the incomplete reporting. The follow-up period of the present data did not necessarily reflect the result of the mass screening test. Moreover, some obstetricians might not regard CH as a birth defect in the narrowest sense . Nevertheless, the objective of this study was to evaluate the incidence rate of CH in multiple births compared to singletons, and not to compare the CH incidence rate across different populations. Therefore, the comparison of CH in multiple births and singletons maybe biased only if there is differential reporting according to plurality, which is not likely to have occurred.To my knowledge, there has been no published study in which the authors directly analyzed the effect of ART on CH. CH incidence rate in developed countries is usually estimated by the data from neonatal screening tests, which are performed after birth. The incidence rate of CH noted in the present study is lower than that noted in many other studies published recently. A neonatal screening test for CH has been mandatory in Japan since 1979. According to the Japanese data on CH, the mean incidence rate of CH was 1/3,600 between 1979 and 2004, and 1/2,000 in 2005, and it then increased slightly to 1/1,800 in 2009 (presented by the Japan Child and Family Research Institute, In the present study, the incidence rate of CH in multiples was about twofold higher than singletons, and even though the difference was not statistically significant, the finding suggests that multiple birth is one of the risk factors of CH. According to the recent population-based case control study by Medda et al. , an incrMany studies ,12-23 reIt has been well established that congenital heart disease is the most frequently occurring additional birth defect in patients with CH ,17,21-25There have been few genetic epidemiologic studies on CH. Family study is a useful tool to show familial aggregation. Intensive family studies on CH were performed by a French group ,28,35 whApart from case reports, the only population-based twin study was performed by Olivieri et al. . AccordiPopulation-based twin study using univariate/multivariate genetic analyses based on the structural equation modeling is the most powerful tool to clarify the genetic/environmental contribution to CH and comorbidity with other birth defects . Record This study has the following limitations, most of which could be attributed to the dataset, based on the fact that individual information was obtained only from the subjects with birth defects and/or CH after ART, not the total ART live births. The first limitation is that the author could not check the reliability of the data directly. This is the essential limitation of secondary data analyses. Second, although the present dataset was from a multi-year nationwide survey, it still did not have sufficiently high statistical power to detect statistical significance. Third, the author could not control for confounding factors that can affect ART and/or CH. Fourth, follow-up after birth was limited to the neonatal period at the latest, and was incomplete . Both CHCH was more frequent in multiples compared to singletons. A higher percentage of additional birth defects was also observed in multiples compared to singletons. The familial aggregation of CH was suggested.CH: Congenital hypothyroidism; ART: Assisted reproductive technology; RR: Relative risk; CI: Confidence interval; RRR: Recurrence risk ratio; PDA: Patent ductus arteriosus; FET: Frozen embryo transfer; IVF: In-vitro fertilization; ICSI: Intracytoplasmic sperm injection.The author declare that I have no competing interests.SO carried out data gathering, analyses, and writing of manuscript.


Emerging evidence has shown that microRNAs (miRNAs) participate in human carcinogenesis as tumor suppressors or oncogenes. Single nucleotide polymorphism (SNP) located in the miRNAs may influence the function of mature miRNAs and then affect the processing of carcinogenesis. It has been suggested that two common SNPs rs2910164 in miR-146a and rs3746444 in miR-499 are associated with susceptibility to hepatocellular carcinoma (HCC). However, published results are inconsistent and inconclusive. To acquire a more precise effect of the association between these polymorphisms and HCC risk, we performed this meta-analysis.We have conducted a search of case-control studies on the associations of SNPs rs2910164 and/or rs3746444 with susceptibility to HCC in PubMed, ScienceDirect, Cochrane Central Register of Controlled Trials, and Chinese National Knowledge Infrastructure databases for the period up to Sep 10th, 2012. A total of 6 studies were identified with 2071 cases and 2350 controls for miR-146a rs2910164 polymorphism, 667 cases and 1006 controls for miR-499 rs3746444 polymorphism. It was found that neither allele frequency nor genotype distribution of the two polymorphisms was associated with risk of HCC in all genetic models. Similarly, subgroup analysis in Asian population showed no associations between the two SNPs and the susceptibility to HCC.This meta-analysis suggests that miR-146a rs2910164 and miR-499 rs3746444 polymorphisms may not be associated with the risk of HCC, especially for Asian population. However, well-designed studies with larger sample size and more detailed data are needed to confirm these conclusions. Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide MicroRNAs (miRNAs) are small non-coding, single-stranded RNA molecules with typical length of 22 nucleotides. MiRNAs are involved in a wide range of physiologic and pathologic processes, including cell differentiation, proliferation, apoptosis and carcinogenesis It is reported that there is association between SNP of rs291016 in miRNA-146a and the susceptibility to HCC Since a single study with a small sample size may not be enough to detect accurate effects of these SNPs on HCC, we performed this meta-analysis to derive more comprehensive and precise estimation of the associations between the SNPs miR-146a rs2910164 and miR-499 rs11614913 and susceptibility to HCC.We carried out a publication search in PubMed, Cochrane Central Register of Controlled Trials, ScienceDirect, and Chinese National Knowledge Infrastructure (CNKI) databases with the following search terms: (“miR-146a” OR “miR-499” OR “rs2910164” OR “rs3746444”) AND (“hepatocellular carcinoma” OR “liver cancer” OR “HCC”) AND (“SNP” OR “mutation” OR “variation” OR “polymorphism”)by two independent investigators . Publication country and publication language were not restricted in our search. We examined reference lists manually to further identify potentially relevant studies, and contacted the corresponding authors of conference abstracts without sufficient data to get additional information by e-mail. If the author had refused to provide the data required in this meta-analysis or we had acquired no reply, the item would be excluded. All the items matching the inclusion criteria were retrieved for further examination and data extraction. Investigators include experts in hepatobiliary, surgery, biologists, epidemiologist and qualified graduate researchers. All of the investigators have received training in literature search, statistics and evidence-based medicine.We set the following criteria for studies recruited in our meta-analysis: (a) evaluated the associations between the SNPs miR-146a rs2910164 and/or miR-499 rs3746444 and susceptibility to HCC, (b) studied on human beings, (c)study designed as case-control, (d) There was sufficient data for the computation of odds ratios and corresponding 95% confidence intervals . (e) If serial studies of the same population from the same group were reported, the latest study would be included. We assessed the methodological qualities of included studies by the description of title, author, year, country, ethnicity, source of sample, gene typing methods, the set of controls and cases, value of Hardy-Weinberg equilibrium (HWE), and factors of risk.Two investigators (Miao Hu and Lianying Zhao) screened titles, abstracts and full texts independently using a standardized screening guide. Data extraction was carried out independently after the concealment of titles, authors, journals, supporting organizations and funds to avoid investigators' bias. After the data abstraction, discrepancies and differences were resolved by consultation and discussion.Characteristics of the enrolled studies were assigned in the structured form , includiThe two investigators (Miao Hu and Lianying Zhao) checked the data extraction results and reached consensus on all of the data extracted. If different results were generated, they would check the data and have a discussion to come to an agreement. Two senior investigators (Surong Hu and Jingting Yang) would be invited to the discussion if disagreement still existed.2 test in controls. The strength of the association between miR-146a rs2910164/miR-499 rs3746444 polymorphism and HCC risk was measured by odds ratios (ORs) with 95% confidence intervals (CIs).Pooled ORs were calculated for allele frequency comparison , additive model ,dominant model , and recessive model respectively. The significance of pooled ORs was determined by Z-test .For each study, the departure of frequencies of miR-146a rs2910164 and miR-499 rs3746444 polymorphisms from expectation under Hardy-Weinberg equilibrium (HWE) was assessed by χStatistical heterogeneity among the studies was checked by chi-square-based Q-test. A P value greater than 0.10 for Q-test indicates no significant heterogeneity existing among studies A total of 136 articles were retrieved after the first search in PubMed, ScienceDirect, Cochrane Central Register of Controlled Trials, and CNKI databases. As shown in The association between miR-146a rs2910164 polymorphism and susceptibility to HCC was analyzed in 6 independent studies with 2,071 cases and 2,350 controls. The result of this meta-analysis is shown in In all the six studies included, five were conducted on Asian, one study on Caucasian. In the ethnicity subgroup analysis, no association was found to be statistically significant on Asian population in any genetic model  .4 studies involving 667 cases and 1,006 controls were evaluated for the association between miR-196a2 rs11614913 polymorphism and HCC risk. Significant statistical heterogeneity was identified in the comparison of allele frequency, AA versus GG, Asian subgroup analysis of dominant model and the recessive model, so that random-effects model was used in these models, Fixed-effects model was used in other models. However, for rs3746444, none of the genetic models produced significant association between rs3746444 polymorphism and HCC risk . The results are summarized in Similarly, subgroup analysis showed no significant association between SNP rs3746444 and susceptibility to HCC in Asian population .Funnel plot and Egger's test were performed to assess the publication bias of the literature . SymmetrWe deleted one single study from the overall pooled analysis each time to check the influence of the removed data set to the pooled ORs. No study was observed to change the homogeneity in heterozygote comparison.The research on microRNAs has provided us a new approach to understand the molecular mechanism of HCC and a new therapy to treat HCC Recently, numerous reports have revealed that SNPs in miRNA-coding region may be involved in the process of hepatocarcinogenesis and contribute to the development of HCC miR-146a rs2910164 is located in the stem region opposite to the mature miR-146a sequence. This C>G polymorphism results in a change from C: U pair to G: U mismatch in the stem structure of miR-146a precursor. Compared with C allele, The G allele gene could lead to more efficient inhibition of target genes including IL-1 receptor-associated kinase 1 (IRAK1), TNF receptor-associated factor 6 (TRAF6) and papillary thyroid carcinoma 1 gene (PTC1) To study the relationship between the miR-146a rs2910164 polymorphism and HCC, we made a meta-analysis including 6 case-control studies with 2071 cases and 2350 controls. Our results did not support a genetic association between rs2910164 and susceptibility to HCC. Neither allele frequency nor genotype distribution was significantly associated with susceptibility to HCC.Considering the potential relationship between the gene polymorphisms in different ethnic groups and the HCC risk, we further made the subgroup analysis on Asian population to explore the association between rs2910164 and the risk of HCC. As a result, no association was observed even in the same ethnic population.Furthermore, deficiency of detailed but important information of both cases and controls such as virus infection, alcohol consumption, age and gender has inhibited us from exploring the relationships between rs2910164 polymorphism and HCC risk more thoroughly. Considering the important biological function of miR-146a in tumorigenesis Another SNP shown to have potential relationship to the risk for HCC is rs3746444 in miR-499. miR-499 was reported to play a role of mediator in a wide spectrum of biological processes, such as cellular senescence, apoptosis, immune response, tumorigenesis and metastasis To study the relationship between the miR-499 rs3746444 polymorphism and HCC, we made a meta-analysis including 4 case-control studies with 667cases and 1006 controls. As a result, our study did not support a genetic association between rs3746444 and susceptibility to HCC. Neither allele frequency nor genotype distribution was significantly associated with susceptibility to HCC. Similar to the miR-146a rs2910164, subgroup analysis on Asian population also showed no association of miR-499 rs3746444 polymorphism with susceptibility to HCC. Sensitivity analysis found no significant influence of any single study on pooled ORs, which indicated the stability of this meta-analysis was acceptable.Firstly, although several meta-analyses have reported associations between these two SNPs and susceptibility to various cancers Secondly, for rs2910164 in microRNA146a, In Wang's study th 2012, several meta-analyses have investigated the relationship between this SNP and risk of cancer Thirdly, for rs3746444 SNP in miRNA-499, by the date of Sep 10In our present meta-analysis, we finished comprehensive literature search in multiple databases as many as possible no matter what the publication languages, dates, ethnicities and other factors that would limit data collection are. Quantitative data synthesis were conducted in allele frequency, additive model, dominant model and recessive model and we found none of the SNPs was associated with the risk of HCC with more powerful evidence compared with previous similar meta-analyses.To the best of our knowledge, this is the first meta-analysis evaluating the potential association between two common polymorphisms rs2910164 in miR-146a and rs3746444 in miRNA-499 and susceptibility to HCC. However, limitations of this meta-analysis should be discussed as they may affect the interpretation of the results.Firstly, the sample size is relatively small although we have searched as many eligible literatures as possible, larger sample size and multiple random-control tests are still needed to detect possible minor effects of miR-146a rs2910164 and rs3746444 in miRNA-499 polymorphisms on susceptibility to HCC.Secondly, lack of available data prevented an adjustment for subgroup factors including alcohol consumption, age, and gender, etc. These factors may become potential determinants to influence the evaluation of the associations between SNPs and susceptibility to HCC by interacting with genetic factors.Thirdly, there was no study on African population in the meta-analysis, and the study on Caucasian is not enough, limitation of the ethnic population enrolled makes the interpretation of the association between the two SNPs and susceptibility to HCC should be more cautious.Finally, it has been generally accepted that HBV is one of the most important factors of HCC In conclusion, this meta-analysis provides more evidence that miR-146a rs2910164 and miR-499 rs3746444 polymorphisms may not be associated with the risk of HCC, especially for Asian population. Well-designed studies with larger sample size and more ethnic groups are of great value to clarify these findings. Moreover, combination genetic variation together with factors such as HBV/HCV infection status, gender, age, and alcohol consumption exposures should also be considered in future.Table S1Checklist.(DOC)Click here for additional data file.Table S2Moose test.(DOCX)Click here for additional data file.


Human serum paraoxonase-1 (PON1) prevents oxidation of low density lipoprotein cholesterol (LDL-C) and hydrolyzes the oxidized form, therefore preventing the development of atherosclerosis. The polymorphisms of PON1 gene are known to affect the PON1 activity and thereby coronary artery disease (CAD) risk. As studies are lacking in North-West Indian Punjabi's, a distinct ethnic group with high incidence of CAD, we determined PON1 activity, genotypes and haplotypes in this population and correlated them with the risk of CAD.350 angiographically proven (≥70% stenosis) CAD patients and 300 healthy controls were investigated. PON1 activity was determined towards paraoxon  and phenylacetate  substrates. In addition, genotyping was carried out by using multiplex PCR, allele specific oligonucleotide –PCR and PCR-RFLP methods and haplotyping was determined by PHASE software. The serum PONase and AREase activities were significantly lower in CAD patients as compared to the controls. All studied polymorphisms except L55M had significant effect on PONase activity. However AREase activity was not affected by them. In a logistic regression model, after adjustment for the conventional risk factors for CAD, QR (OR: 2.73 (1.57–4.72)) and RR ) genotypes of Q192R polymorphism and GG (OR: 2.07 (1.02–4.21)) genotype of −162A/G polymorphism had significantly higher CAD risk. Haplotypes L-T-G-Q-C (OR: 3.25 (1.72–6.16)) and L-T-G-R-G (OR: 2.82 (1.01–7.80)) were also significantly associated with CAD.In conclusion this study shows that CAD patients had lower PONase and AREase activities as compared to the controls. The coding Q192R polymorphism, promoter −162A/G polymorphism and L-T-G-Q-C and L-T-G-R-G haplotypes are all independently associated with CAD. They are thought to be progeny of Indo – Scythian, Indo European and Indo – Aryan stocks In the CAD group, 180 patients had a single vessel involved whereas 190 had double and 52 had triple vessel disease. Compared to the controls, CAD group had greater proportion of men, alcohol consumers and were significantly older, had higher BMI and lower serum HDL-C levels (p = 0.0001) . HoweverThe CAD patients had significantly lower serum PONase (113.0 nmol/min/ml vs. 178.0 nmol/min/ml) and AREase activities (74.6 µmol/min/ml vs. 82.7 µmol/min/ml) (P<0.0001) as compared to the controls . The HDLL55M polymorphism had no effect on serum PONase and AREase activities in both the groups. However when comparison was carried out between groups within genotypes, both activities was significantly lower in LL, LM and MM genotypes in CAD patients as compared to the controls . PONase HDL-C was significantly lower (P<0.05) in QQ (Q192R), and GG (−162A/G) genotypes and TG was significantly higher (P<0.05) in GG (−909G/C) and RR (Q192R) genotypes in patients as compared to controls. No significant difference was observed for LDL-C and TC in any of the group (data not shown).The genotype and allele frequency distribution of coding Q192R and L55M polymorphisms of CAD patients and controls is shown in Association between Q192R polymorphism and CAD was followed up with multiple logistic regression to determine if the association was independent of other known risk factors for CAD, such as alcohol consumption, HDL-C levels, age, and sex. In the regression analysis, age, sex, alcohol consumption, HDL-C levels and BMI were associated with CAD. The association between Q192R polymorphism and CAD persisted in this analysis. The ORs were 2.73 (95% CI: 1.57–4.72) and 16.24 (95% CI: 6.41–41.14) for QR heterozygote and RR homozygote respectively .In promoter −909G/C polymorphism, GC and CC genotypes were significantly associated with CAD (P = 0.0001) with ORs being 1.80 (95% CI: 1.24–2.59) and 3.03 (95% CI: 1.96–4.68) respectively  as compaHowever after multiple logistic regression analysis in which cardiovascular risk factors were adjusted, the association was observed only with −162A/G polymorphism and CAD with OR being 2.07 (95% CI: 1.02–4.21) for GG homozygotes . No signLD values were generated to look for association among the five polymorphisms. No significant LD was observed among the PON1 SNPs with D' value ranging from 0.03 to 0.25 .On haplotype analysis we observed 32 different combinations of these coding and promoter polymorphisms . The freThe susceptibility of developing CAD in Asian Indians is 3–4 times higher than Caucasians, 6-times higher than Chinese, and 20-times higher than Japanese Previous studies that have investigated the relationship between PON1- Q192R polymorphism and CAD have produced inconsistent results. Wheeler et al. We found lower PON1 activity towards paraoxon (PONase) substrate in CAD patients as compared to the controls, with results being similar to prospective epidemiological Caperhilly PON1 activity is not affected towards phenylacetate (AREase) substrate across Q192R polymorphism Fewer studies have been conducted into the relationship between the PON-L55M polymorphism and CAD but again with inconsistent results GG<AG<AA genotypes within the −162 A/G polymorphism, CC<CG<GG genotypes in −909G/C polymorphism and TT<CT<CC genotypes within −108C/T polymorphisms in patients and controls. These observations are similar to other studies CGGGGG consensus sequence, which is the binding site for the sp1 transcription factor To the best of our knowledge, not much information is available in literature in terms of PON1 promoter polymorphisms, PON1 activity and CAD risk. We found that PONase activity was significantly higher in wild AA (−162 site), GG (−909 site) and CC (−108C/T site) homozygotes. It increased in the order of the AREase activity was not affected by any of the promoter polymorphisms in any group. These results indicate that AREase activity is a better enzymatic test for examining the association between PON1 activity and disease, especially when genotyping cannot be done. In some studies linkage disequilibrium has been observed between PON1 polymorphism at position 55 with that at position 192 Determination of haplotypes is gaining attention because multiple linked SNPs have the potential to provide significantly more power to genetic analysis than individual SNPs In a study of the Hutterite Brethren, a North American population isolated by religious belief, the PON1 genotype was significantly associated with variation in the concentration of HDL-C, LDL-C, TG and apo-B In conclusion CAD patients had lower PONase and AREase activities as compared to the controls. The coding Q192R, promoter −162A/G polymorphisms and L-T-G-Q-C and L-T-G-R-G haplotypes are all independently associated with CAD.The present study has several strengths and limitations that need to be addressed briefly. The strengths include the use of a pure Punjabi population from the North-West region of India to eliminate false positive results due to population stratification. Angiographical profiles of all subjects were evaluated. Regarding limitations, we could not stop statins in CAD patients due to ethical reasons. As statins are known to influence PON1 activity Between August 2007 and September 2009, 350 North-West Indian Punjabi's with angiographically proven CAD at Nehru hospital , Postgraduate Institute of Medical Education and Research, Chandigarh (India) were included in the study. Patients with diabetes mellitus, cirrhosis of liver, COPD, HIV infection, malnutrition and acute myocardial infarction were excluded from the study. We also excluded those patients who had undergone a recent coronary intervention i.e. angioplasty or bypass. The disease severity was determined by counting the number of affected three major epicardial coronary arteries with significant stenosis (≥70%). The angiogram was assessed by at least two cardiologists who were unaware of the patient's status before inclusion in the study. In addition, relevant history was recorded regarding risk factors for CAD such as smoking, alcohol consumption, presence or absence of CAD among first degree relatives (parents and siblings) and the age of onset and medical treatment. Due to ethical reasons it was not possible to stop treatment including statins in patients. The dose of atorvastatin varied from 20–80 mg/day with majority on 40 mg. In the control group, 300 North-West Indian Punjabi's who were attending a general medical clinic for health check up and were found to be healthy were included.All the subjects were provided with the study protocol and an informed written consent was obtained. The study was approved by the institute  ethics committee.Venous blood sample was collected from both controls and CAD patients between 8.30 and 9.30 AM after an overnight fast. The serum was isolated by low-speed centrifugation and enzymatic assays were carried out. The lipid profile  was measured by enzymatic method using autoanalyzer (Hitachi modular P) and Roche diagnostic kits. The PON1 activity and level were measured by using paraoxon and phenylacetate  as substrates respectively with activity was measured by a modification of method described by Mackness et al. The DNA was extracted from the blood cells by method described by Daly et al. Statistical analysis was carried out using SPSS (Version 17.0) software. All the study variables are shown as mean ± standard deviation or median (range) depending on the shape of the distribution curve. Student's t-test or Mann-Whitney U test was used to compare the data between controls and CAD patients. The demographic parameters were correlated with CAD by univariate logistic regression analysis, and followed up with multiple logistic regression analysis for significant variables. A multiple linear regression analysis was undertaken to determine the independence of PON1 activity from other variables.2 test was used for comparison between groups. The genotypic association with CAD and their odds ratio (OR) with 95% confidence interval was estimated by binary logistic regression, where homozygous wild type was kept as a reference. The effect of genetic variants on CAD susceptibility was determined by multiple logistic regression analysis after adjusting the conventional risk factors. Power of the sample size was calculated using the PAWE software http://www.broad.mit.edu/mpg/haploview/contact.php) and was expressed in terms of D'. The haplotype frequency was determined by PHASE (Version 2.1) software (http://stephenslab.uchicago.edu/software.html). Fisher's exact test was used to find differences in haplotype frequency between CAD patients and controls. A p value≤0.05 was considered significant.The genotype and allele frequencies for each polymorphism were stratified for homozygote wild, heterozygote and homozygote variant type of the respective allelic variant and Pearson's χTable S1The haplotypes frequency distribution in Controls and CAD patients for coding  and promoter  SNPs of PON1 gene.(DOC)Click here for additional data file.Table S2PCR conditions used in genotyping of promoter SNPs of PON1 gene.(DOC)Click here for additional data file.Text S1(DOCX)Click here for additional data file.


Allelic polymorphism in codon 72 of the p53 tumor suppressor gene causes imbalance of p53 protein expression. Earlier studies have shown association between allelic polymorphism in codon 72 of the p53 gene with risk of ovary cancer (OC); however the results are inconclusive and conflicting. Therefore, we performed this meta-analysis to investigate the relation between p53 codon 72 Arg>Pro polymorphism and overall OC susceptibility.We searched all eligible published studies based on the association between codon 72 of the p53 Arg>Pro polymorphism and risk of OC. Data were pooled together from individual studies and meta-analysis was performed. Pooled odds ratios (ORs) and 95% CI were calculated for allele contrast, homozygous, heterozygous, dominant and recessive genetic models.A total of twelve studies comprising of 993 OC cases and 1264 healthy controls were included in this meta-analysis. Overall, no significant association was detected for Pro allele carrier , homozygous , heterozygous , dominant , and recessive  genetic models, respectively. Also, in the stratified analysis by ethnicity, no significant association of this polymorphism with risk of OC was found in the Caucasian population.This meta-analysis suggested that codon 72 of the p53 Arg>Pro polymorphism may not significantly contribute in ovary cancer susceptibility. However, future large studies with gene-gene and gene-environment interactions are needed to validate these findings. Ovary cancer (OC) is the most common carcinoma among females with poor prognosis. It is the sixth leading cause of death among gynecological malignancies in females worldwide The p53 tumor suppressor gene (TP53 at 17p13), recognized as “the guardian of the genome”, plays a significant role in the cell cycle arrest, senescence, DNA damage repair, regulates the cell cycle and requires loss of function mutations for tumor formation The p53 protein functions by interfering with central regulators of hypoxia which mediate angiogenesis, and eventually inhibit production of pro-angiogenic factors and endogenous angiogenesis inhibitors Several polymorphisms have been detected in both coding and non-coding region of this gene Considering the functional significance of p53 gene in carcinogenesis, it is speculated that codon 72 Arg>Pro polymorphism may be a potential susceptibility factor for OC. Lately, several epidemiological case-control studies have evaluated the association between p53codon 72 Arg>Pro polymorphisms and OC risk We carried out a PubMed (Medline), EMBASE and Google Scholar web database search covering all research articles published with a combination of the following key words: ‘p53 gene (polymorphism OR mutation OR variant) AND ovarian carcinoma or ovary cancer, tumor susceptibility (last updated on November 2013). All the searched studies were retrieved and their reference lists were checked as well for other relevant studies. When, more than one of the same population was included in several publications, only the most recent or complete study was included in this meta-analysis. Since, this is a meta-analysis of published articles based on the association of p53 codon 72 Arg>Pro polymorphism and OC risk, so ethical approval was not required for this study.In order to minimize heterogeneity and ease the appropriate interpretation of this study, published articles included in the current meta-analysis had to meet all the following inclusion criteria: a) studies should have a cross-sectional, case-control or cohort design, b) must evaluated the association between p53 codon 72 Arg>Pro polymorphism and OC risk, c) recruited pathologically or histologically confirmed OC patients and healthy controls, d) have available genotype frequency in case and control, e) and published in English language. Also, when the case-control study was included by more than one research article using the same case series, we selected the study that included the largest number of individuals. On the other side, the major reasons for study exclusion were, overlapping of the data, case-only studies, review articles, and genotype frequencies or numbers are not reported. The study selection procedure has been shown in the form of flow-diagram as For each retrieved research publication, the methodological quality assessment and data extraction were independently abstracted in duplicate using a standard protocol by two independent investigators. Data-collection form was used to guarantee the accuracy of the collected data by stringently following the inclusion-exclusion criteria mentioned above. The main characteristics abstracted from the retrieved studies included the name of the first author, publication year, the country of origin, the number of cases and controls, study type, and genotype frequencies. Cases associated with disagreement on any item of the data from the collected research studies were fully debated with investigators to achieve a final consensus.2 statistics was employed to quantify inter-study variability and larger values suggested an increasing degree of heterogeneity http://www.meta-analysis.com/pages/comparisons.html. The Comprehensive Meta-Analysis (CMA) Version 2 software program  was chosen and utilized to perform all statistical analysis involved in this study.In order to examine the relationship between p53 codon 72 Arg>Pro polymorphism and OC risk, pooled ORs and their corresponding 95% CIs were estimated. Heterogeneity assumption was examined by the chi-square-based Q-test According to our selection (inclusion-exclusion) criteria, a total of twelve research articles were finally included through literature search from the PubMed (Medline), EMBASE and Google Scholar web databases in this meta-analysis. All retrieved research publications were examined carefully by reading the titles and abstracts, and the full texts for the potentially relevant research articles were further checked for their aptness for the current meta-analysis. Studies either showing p53 codon 72 Arg>Pro polymorphism to predict survival in OC patients or considering p53 variants as an indicators for response to therapy were excluded straightaway. Similarly, studies investigating the levels of p53 mRNA or protein expression or relevant review articles were also excluded. In the present meta-analysis, only case-control or cohort design studies having frequency of all three genotypes were included. Besides the database search, the reference lists present in the retrieved articles were also checked for other potential research publications . DistribBegg's funnel plot and Egger's test were carried out to evaluate the publication bias among the selected studies for the meta-analysis. The appearance of the shape of funnel plots was seemed to be symmetrical in all the genetic models. The Egger's test was performed to provide the statistical evidence of funnel plot. The results showed lack of publication bias among all comparison models .2 statistics were employed and heterogeneity was noticed in all the five genetic models. Therefore, random effects model was applied to synthesize the data , homozygous , heterozygous , recessive  and dominant model  .We have analyzed only Caucasian population by study design and participants. This meta-analysis included eight studies (717 cases and 797 controls), Heterogeneity was observed in all genetic models; thus, random effect model was applied to analyze the data. In subgroup analysis publication bias did not exist . We did The p53 tumor suppressor gene, mutated in many human cancer types, suggests its key role in the host's defense against malignancy Despite remarkable progress in mechanistic understanding of p53 structure and function, the contribution of specific p53 gene polymorphisms to OC risk remains equivocal and proven to be extremely complex biomarkers. Interest in the genetic susceptibility to OC has led to an emerging trend to the study of polymorphisms of genes involved in OC risk. Due to the different roles of p53 gene in human genome, it has been hypothesized that codon 72 Arg>Pro polymorphism is associated with risk of OC. As a result, a large number of studies have been performed to evaluate the association between p53 codon 72 Arg>Pro polymorphism and risk of OC, but the results from different published studies lacks consensus. Inconsistency in results from these studies can be attributed to low statistical power to evaluate the overall effect of the p53 codon 72 Arg>Pro polymorphism with OC risk. The answer to this limitation is a meta-analysis, a powerful tool for investigating the risk factors associated with genetic diseases, which employs quantitative technique to pool the data from individual studies where individual sample sizes are small with lower statistical power, and provides reliable conclusion The overall pooled results of this meta-analysis revealed that p53 codon 72 Arg>Pro polymorphism did not influence an increased or decreased risk of OC in all the five genetic models as per the eligible studies when compared with wild type allele. Even in the stratified analysis by ethnicity, no statistically significant relationship between p53 codon 72 Arg>Pro genotype and OC risk was detected in Caucasian population. One possible explanation is that several other SNPs have been reported in the p53 gene and previous research showed that various other SNPs are related to the susceptibility to OC. It is possible that the analyzed variant does not act as primary susceptibility polymorphism and may be inhibiting p53 function by linking with other functional polymorphism alleles found in linkage disequilibrium (LD). Our results are in agreement with Shen et al. Heterogeneity between studies is very common in the genetic association studies of meta-analysis. In the present meta-analysis we found inter-study heterogeneity in overall analysis. There are several factors responsible for such heterogeneity, i.e., the genetic backgrounds for cases and controls, diverse genotype distribution of codon 72 Arg>Pro in different ethnic groups and uneven selection criteria for the cases and controls in different studies.Despite the important findings from our current analysis, we still have to acknowledge some limitations of this study. First, we only included studies published in English language, abstracted and indexed by the selected electronic databases for the data analysis; it is possible that some pertinent reports published in other languages and indexed in other electronic databases may have missed. Second, the result of this meta-analysis was based on unadjusted ORs because not all eligible studies stated adjusted ORs. Third, the role of gene-environment interactions were not considered which may affect the risk of OC. Also, it is worthwhile to mention several strengths of our study. First, we have included significantly more number of cases and controls comparison to the previous meta-analysis study by using effective and efficient search strategy to increase the statistical power of the analysis. Second, the quality of the case-control studies included in the present pooling analysis was satisfactory and met with the pre-set inclusion criteria.In conclusion, a meta-analysis is a rational approach of data-analysis which pools both statistically significant and non-significant findings from individual studies to improve the statistical performance by increasing the sample size. Our meta-analysis demonstrates that p53 codon 72 Arg>Pro polymorphism might not significantly modulate the OC risk. However, future well designed large studies, particularly stratified by gene-gene and gene-environment interactions might be necessary to clarify the possible role of the p53 codon 72 Arg>Pro polymorphism in the susceptibility to OC.Figure S1PRISMA 2009 Flow Diagram. Showing identification and selection of studies for the meta-analysis.(TIF)Click here for additional data file.Checklist S1PRISMA 2009 Checklist.(DOC)Click here for additional data file.


A large number of studies have investigated whether polymorphisms in the Toll-like receptor (TLR) genes are implicated in susceptibility to tuberculosis (TB) in different populations. However, the results are inconsistent and inconclusive.TLR1 G1805T, TLR2 T597C, T1350C, G2258A, and TLR6 C745T polymorphisms and TB risk was carried out by comparison using different genetic models.A literature search was conducted using the PubMed, EMBASE, Medline (Ovid), ISI Web of Knowledge and Chinese National Knowledge Infrastructure (CNKI). A meta-analysis on the associations between the TLR2 2258AA  and TLR6 745TT  polymorphisms and TB risk. In the subgroup analysis by ethnicity, Africans and American Hispanic subjects with the TLR1 1805T allele had an increased susceptibility, whereas Asian and European subjects with the TLR2 2258A allele had an increased susceptibility to TB.In total, 16 studies from 14 articles were included in this review. In meta-analysis, significant associations were observed between the TLR2 G2258A is associated with increased TB risk, especially in Asians and Europeans. TLR1 G1805T is associated with increased TB in Africans and American Hispanics. TLR6 C745T is associated with decreased TB risk. Our systematic review and meta-analysis reported an interesting preliminary conclusion, but this must be validated by future large-scale and functional studies in different populations.The meta-analysis indicated that  Tuberculosis (TB) is a contagious and potentially fatal disease caused by various strains of mycobacteria, usually Mycobacterium tuberculosis (Mtb) in humans. It can infect almost any part of the body, but manifests mainly as an infection of the lungs and kills more people each year than any other single infectious disease. It is estimated by WHO that about one-third of the current global population is infected asymptomatically with Mtb, of whom 5–10% will develop clinical disease during their lifetime TLR4 Asp299Gly, Thr399Ile, and TIRAP Ser180Leu polymorphisms, which are the most studied SNPs in these two genes TLR genes and their association with TB. Among them, TLR1 G1805T, TLR2 T597C, T1350C, G2258A, TLR6 C745T are most studied as potential risk factors for TB. TLR2 T597C, T1350C and TLR6 C745T are in the extracellular domains, TLR1 G1850T is in the trans-membrane (TM) domain and TLR2 G2258A is in the Toll-Interleukin-1 receptor (TIR) domain.Toll-like receptors (TLRs) can recognize pathogen-associated molecular patterns (PAMPs) of Mtb and initiate signaling pathways that lead to the activation of the innate immune response, cytokines and formation of the adaptive immune response st January 2013 to summarize the associations between these polymorphisms and TB susceptibility.Many studies have reported the association of these polymorphisms with TB; however, the results are inconsistent and inconclusive due to limited sample sizes and different study populations. Therefore, in this article, we chose these polymorphisms as our candidates and performed a systematic review and meta-analysis, based on literature identification until 1st January 2013. The key words were: ‘Mycobacterium tuberculosis’ or ‘tuberculosis’ in combination with ‘polymorphism’ or ‘variant’ or ‘genotype’ or ‘allele’ or ‘mutation’, and in combination with ‘toll’ or ‘TLR’ or ‘Toll-like receptor’ or ‘Toll like receptor’. The search results were limited to English and Chinese language articles. All retrieved titles and abstracts were examined for relevant studies on the association between TB and TLR gene polymorphisms. Studies were selected if they met the following criteria: 1) case-control studies of unrelated individuals; 2) evaluation of polymorphisms TLR1 G1805T, TLR2 T597C, T1350C, G2258A, TLR6 C745T and TB susceptibility; 3) genotype distribution in both cases and controls were available. Exclusion criteria were: 1) study design based on family or sibling pairs; 2) genotype frequencies not reported; and 3) data from reviews and abstracts. Additional studies were also identified by hand searching reference lists of original studies and review articles including meta-analysis. The PRISMA Checklist in We searched the PubMed, EMBASE, Medline (Ovid), ISI Web of Knowledge and Chinese National Knowledge Infrastructure (CNKI) to identify studies of the association between TB susceptibility and TLRs polymorphisms until 1For all studies, we extracted the following data from original publications: first author and year of publication; distribution of genotypes for each polymorphism among cases and controls; characteristics of the study design and the study population .2 test P-value was less than 0.05. Fixed-effects models were adopted when P-value is more than 0.05; otherwise random-effects models were used. The funnel plot, Begg’s test and Egger’s test were used to evaluate the publication bias Hardy-Weinberg Equilibrium (HWE) was examined in controls by asymptotic Pearson’s Chi-square test for each polymorphism in each study. The association between polymorphism and TB was estimated by means of odds ratios (OR) and corresponding 95% confidence intervals (CI) comparing cases to controls. Between-study heterogeneity was tested by the Q test and ITLR1 G1805T, seven studies for TLR2 T597C, six studies for TLR2 T1350C, eleven studies for TLR2 G2258A, and four studies for TLR6 C745T. As shown in A total of 527 articles were achieved by literature search. As shown in Six case-control studies (1684 cases and 1216 controls) on the relationship between the G1805A polymorphisms and the risk of TB were included in the meta-analysis.2 value was 40.05, with 5 degrees of freedom (df) and P<0.001 in a random effect model. I2, another index of the test of heterogeneity, was 88%, suggesting a high heterogeneity. We therefore chose the random-effect model to synthesize the data. The overall OR for the T vs. G alleles was 1.17  were included in the meta-analysis on the relationship between the T597C polymorphism and the risk of TB.2 value was 15.14, with 6 df and P = 0.02 using the random-effect model, while I2 was 60, suggesting a modest heterogeneity. We therefore chose the random-effect model to synthesize the data. The overall OR for the C vs. T alleles was 0.98 (95% CI 0.83–1.15) and the test for overall effect, Z value, was 0.24 (P = 0.81), indicating no association between this polymorphism and TB. The publication bias by Begg’s test showed no significant publication bias (P>0.05). Comparisons for four other genetic models were also performed, and no associations were found either (As shown in d either .Six case-control studies (1379 cases and 1309 controls) were included in the meta-analysis to determine the relationship between the T1350C polymorphism and the risk of TB.2 value was 8.79, with 5 df and P = 0.12 using the fixed-effect model, while I2 was 43%, suggesting limited heterogeneity (P = 0.59). Comparisons for four other genetic models were also performed, and no associations were found (P<0.05), indicating that there might be some publication bias in these studies.As shown in ogeneity . The overe found . The pubTLR2 G2258A were included in the meta-analysis.Eleven case-control studies (2823 cases and 2254 controls) on the relationship between the 2 value was 20.64, with 9 df and P = 0.01 in a random-effect model and I2 was 56%, suggesting moderate heterogeneity. The overall OR for the A vs. G alleles was 1.78 (95% CI 0.99–3.20) and the test for overall effect, Z value, was 1.92.  using the random effect model, indicating an association of the AA genotype with TB risk. The Begg’s test did not show publication bias in the studies . Significantly increased risks were found among Asians  were included in the meta-analysis on the relationship between the 2 value was 12.64, with 3 df and P = 0.005 using the random-effect model, while I2 was 76, suggesting a high heterogeneity. The overall OR for the C vs. T alleles was 0.66 (95% CI 0.44–0.99) and the test for overall effect, Z value, was 1.34 , also indicating a decreased TB risk in subject with the TT genotype. The publication bias by Begg’s test showed no significant publication bias  and TB risk reported until 1st January 2013. The meta-analysis revealed an association between the TLR2 2258AA genotype and TLR6 745TT and TB risk. At the same time, different genetic models and ethnicity subgroups comparisons were also performed in TLR1 G1805T, TLR2 G2258A polymorphisms. TLR1 1805T allele was associated with TB in Africans and American Hispanics, and TLR2 2258A allele was associated with TB in Asians and Europeans. However, the TLR2 T597C, TLR2 T1350C polymorphism did not show significant association with TB.In this study, we performed a meta-analysis to assess the association between five extensively studied polymorphisms , but other polymorphisms in the TLR2 did not associate with TB. The subgroup analysis by ethnicity showed that G2258A was associated with an increased risk of TB in Asian and European populations. The reasons for this difference of association in different populations were unclear, but the differences in the frequencies of the polymorphisms among different races might contribute to this heterogeneity. In addition, numerous polymorphisms outside the TLR2 gene can contribute to the host susceptibility to TB. These polymorphisms might differ greatly in various ethnic groups with different evolutionary backgrounds. These differences, along with gene-gene interactions and environmental and cultural factors, and even the variations in Mycobacterium strains, made understanding the observed differences between ethnic groups even more complicated.The G2258A polymorphism in TLR2 was first identified by mutation screen assay in healthy study subjects, which cause the 753 amino acid substitution from Arg to Gln in the conserved TIR domain  [37]. InTLR1 T1805G is a common non-synonymous polymorphisms located in the trans-membrane domain of TLR1 molecule .(TIF)Click here for additional data file.Figure S2Funnel plot analysis to detect publication bias for association of TLR2 G2258A and TB risk in allele comparison (A vs. G).(TIF)Click here for additional data file.Figure S3Funnel plot analysis to detect publication bias for association of TLR6 C745T and TB risk in allele comparison (T vs. C).(TIF)Click here for additional data file.Table S1PRISMA 2009 checklist.(DOC)Click here for additional data file.Table S2Exclusion criteria for excluded studies.(DOC)Click here for additional data file.


Women treated for Hodgkin’s lymphoma (HL) have an elevated risk of developing second breast cancer (SBC) compared with the general population. We planned this meta-analysis to quantify the long-term risk of SBC and analyze the contributing risk factors among HL survivors.According to predefined selection criteria, literature search identified 34 studies that were included in the analyses.I2 = 96%), with a median absolute excess rate of 22.9 per 10,000 person-years. The RR was found inversely related to age at diagnosis of HL with the highest rate , occurred in young patients (≤ 15 years old), where the RR in older women (≥ 40 years old) was not significant . Analysis of RR by 5-year increments since the treatment of HL showed that the risk was highest after 15–19 years of latency . Analysis of the effect of treatment modalities showed that the RR rates were , , and , for radiotherapy (RT) only, combined RT and chemotherapy (CT), and CT only, respectively. To investigate the demonstrated heterogeneity, meta-regression analysis was performed when feasible. In most such analyses, the natural logarithm of RR was inversely associated with age at HL diagnosis.After eliminating overlapping or duplicate data, 957 incidences of SBC were encountered in 24,505 females with HL over a median follow-up of 14.9 years. The medians: age at the diagnosis of HL, age at diagnosis of SBC, and latency since HL treatment to the development of SBC were 23.7, 35.0, and 17.7 years, respectively. The pooled relative risk (RR) of SBC was 8.23 (95% CI, 5.43-12.47, We conclude that, the current meta-analysis provided the most recent comprehensive estimate of the risk of SBC in a broad-range of HL survivors. Younger age at diagnosis proved to be a dominant risk factor. The obtained results would serve providing breast cancer screening recommendations for HL survivors. Hodgkin’s lymphoma (HL) became a curable disease by radiation therapy (RT) and/or combination chemotherapy (CT) since the early 70s -3. Long-However, increased risk of second cancer following effective treatment of HL has long been reported . More reTwo pertinent meta-analyses have been published and they have addressed different questions. The first meta-analysis was published in 2006 and examined all second malignancy risk associated with HL treatment in 31 randomized trials and it included 65 incidence of SBC . In the Research on the late consequences of HL has often been limited by the size and composition of the study populations and by the duration and completeness of patient follow-up. To the best of our knowledge, there is no recently published meta-analysis intended to examine the risk of SBC in a broad range of ages at HL diagnosis, various follow-up periods, and subsequent to different therapeutic modalities. Also not precisely known, is the effect of other contributing risk factors. The lack of such data has prompted the current meta-analysis.Between January 1966 and October 2011, we identified studies of interest by first conducting an electronic literature search of the databases MEDLINE, EMBASE, and the Cochrane Library. We also searched for relevant abstracts in the annual conference proceedings between January 1984 to October 2011 for the American Society of Clinical Oncology, European Society for Medical Oncology, and the San Antonio Breast Cancer Symposium. All ages of HL patients were eligible for inclusion.We used exploded Medical Subject Heading terms or key words terms ‘lymphoma’, ‘Hodgkin’, ‘Hodgkin’s disease’ and ‘Hodgkin’s lymphoma’. The terms were combined with ‘neoplasm, second neoplasm, second primary’ using the Boolean operator ‘and’. Search results were also filtered against the terms ‘breast, breast cancer, breast neoplasm). In the second step, these keywords were combined using the Boolean operator ‘and’ with ‘standardized incidence ratio’, ‘relative risk’, and ‘observed to expected’. In addition, we manually reviewed the reference lists of relevant studies to identify additional pertinent published articles.We included studies that met each of following criteria: (i) published in English language between January 1985 and October 2011; (ii) included naive patients at any age and with any stage of HL; (iii) investigated the risk for second malignant neoplasms (SMNs) in HL survivors; (iv) reported relative risk (RR) and/or specified as standardized incidence ratios (SIR) or data allowing such outcomes to be derived; and (v) published as original articles .When two or more references reported duplicate data, we only included in the analysis the most recent data, studies with the longer follow-up, or the most relevant studies. We excluded studies that mainly addressed the clinical characteristics of SBC. We also excluded studies that mainly intended to evaluate the potential benefits and harms associated with breast cancer surveillance among women with HL. Case–control designs, i.e. HL patients who developed BC compared with patients who did not were excluded.Two authors  independently inspected each reference title identified by the search and applied the inclusion criteria. For possibly relevant articles and in cases of disagreement between reviewers, the full article was obtained and inspected independently by the five authors. The data intended for extraction were discussed, and decisions were documented. We used the STROBE  reporting criteria to assess the quality of studies included in the meta-analysis . Any sig1a/2- ). When appropriate, we also used the built-in calculator of the Review Manager Software  to compute missing data. When studies showed that the observed number of cases was zero, we simply added 1 to both the observed and the expected number of cases to allow computation of an estimate of the lnRR and its associated SE [Standardized Excel sheet was used for each study that fulfilled the inclusion criteria. Extracted data included paper characteristics , study design, number of HL patients, mean/median age of patients, mean/median duration of follow-up, therapy details, number of observed and expected SBC cases, and RR or standardized-incidence rate (SIR) with corresponding 95% confidence interval (CI). The ratio of observed to expected numbers of cancers, SIR (referred to in the text as RR) was then used or calculated with likelihood-based 95% CI from Poisson models . Where niated SE .The primary outcome was the overall pooled RR of incidence of SBC among women survivors of HD. The secondary end points were RR vs. various variables: source of data, age at diagnosis of HD, length of follow-up, treatment modalities, and any additional relevant risk factors. RT in this meta-analysis is referred to supra-diaphragmatic irradiation with or without other radiation fields.x2 test of heterogeneity and the I2 statistic of inconsistency [x2 P value less than .1 or an I2 statistic greater than 50%. The estimates of RR, together with associated 95% CI, were obtained using the DerSimonian and Laird random-effects model [We assessed heterogeneity of the studies’ results by inspecting graphical presentations and by calculating an sistency ,20. Statts model . Meta-reSubgroup analyses were performed to assess potential contributions of various clinical variables to outcomes. A funnel plot estimating the precision of trials (plots of logarithm of the RR against the sample size) was examined for asymmetry to estimate publication bias . PublicaAll statistical tests were two-sided. RR was estimated according to the inverse of variance method with the use of Review Manager Software v5.1.4.We identified 1,647 potentially relevant articles .The remaining 34 studies were included in the meta-analysis. The funnel plot of 25 non-overlapping studies showed mild asymmetry, however, the Egger linear regression test was not significant  of SBC incidence per 10,000 person-years of observation was 22.9 excess cases . Few studies reported the cumulative incidence of SBC after 20 and 30 years of follow-up .I2 statistic = 96%; p <0.0001). The random-effects model was computed instead and it showed that patients with HL have an almost 9-fold increase in the risk of SBC . We performed meta-regression analysis to determine to what extent the heterogeneity is explained by the effects of study size, age at HL diagnosis, and the latency since the completion of HL treatment. The univariate analysis showed significant inverse association between lnRR and age at diagnosis and a positive relation to latency since HL treatment. With multivariate analysis  was explored by meta-regression analysis, however, none of variables tested was found associated with lnRR in the univariate analysis (data not shown).In Figure I2 = 79%), we performed a meta-regression analysis, however, none of the explanatory variables was found significant (data not shown).The excess risk of SBC as a function of age at diagnosis of HL was also explored. Figure I2 = 41%). Of note, the RR of SBC for women who developed HL above the age of 40 years was not significant .Several studies reported the RR at certain cut points of age at diagnosis and it also showed an inverse relation between risk and age . By and large, the analysis demonstrated an increasing RR by increased duration of follow-up latency reaching the highest after 15–19 years . While there was a decrease in RR noted after 20–24 years of follow-up, further rise occurred after 25–29 years. The latter rise may be attributed to the RR reported by De Bruin et al. [We performed analysis of RR by 5-year increments since the treatment of HL . RT used as the sole therapeutic modality was associated with an almost 5-fold increase in risk , I2 = 74%) and even higher rate  when RT was used for patients ≤ 30 years of age [Figure s of age ,36,41. Ts of age ,54, repos of age , 1 patieI2 = 91%). Nevertheless, adding alkylating CT to RT did not abate SBC risk , while the combination of RT and non-alkylating CT caused a non-significant effect  (data not shown). Noteworthy, only a few studies have provided enough data about the nature of CT offered. The current meta-analysis showed that the use of CT only was not associated with significant risk .Figure To explore the heterogeneity of the RR vs. therapeutic modalities, we performed a univariate analysis that showed an inverse association between lnRR and age at HL diagnosis, positive interaction with latency, and no significant effect for study size. The multivariate meta-regression analysis, however, only showed that younger age at diagnosis retained a significant independent risk Table .Comparing HL patients diagnosed from 1960s to the early 1970s, late 1970s to early 1980s, and more recent years the reported RRs were 3.7, 5.9, and 10.7, respectively ,48,51,55Table HL has been a successful model for the development of effective treatment approach in clinical oncology. Long-term survivors of that disease have also allowed better recognition and understanding of the late effects of therapy. In a large cohort of 25,305 women with HL, and with 957 incidences of SBC reported from North American and European institutions, the current meta-analysis quantified the risk of SBC. The overall pooled analysis showed that there was an approximate 9-fold increase in the risk of SBC incidence (pooled RR = 8.23), and AER of 23 patients per 10,000 person-years.Patients included in this meta-analysis developed HD at a median age of 23.7 years. There has been incongruity about the influence of younger age at HL diagnosis and the higher risk of SBC, where some studies have failed to prove that effect ,42. The RT used as the sole therapeutic modality was associated with a 5-fold (RR = 4.70) increase in risk and a 14-fold (RR = 14.08) increase among young (≤ 30 years of age). Almost all SBC arose within or at the margin of the RT field. The RR of combined RT and any CT was slightly higher than that associated with RT only 5.65 vs. 4.70). An even higher, was the RR associated with the combination of RT and alkylating CT (RR = 6.59), thus, the potential protective effect of gonado-toxic alkylating CT was not demonstrated. Several studies showed an inverse association between the use of alkylating CT in HL and SBC risk ,65, howe vs. 4.70Analysis of potential additional risk factors was limited due to lack of sufficient data and/or inability to compute missing information, therefore, cautions should be exerted in interpreting results. Two studies ,54, repoThe reason for failing to show a convincing evidence of RT dose–response effect associated with SBC risk is at best divisive. There is evidence for a strongly linear radiation dose response, but only in the lower dose range (up to 5 or 10 Gy) ,68. It hAlthough new RT techniques and treatment strategies have the potential to reduce the future burden of late effects, nevertheless, we have shown that an even higher risk was reported in more recent years suggesting that there remains a significant cohort at an increased risk of SBC.Pelvic RT was found to be associated with a protective effect as reported by De Bruin et al. . The samOnly one study examined the receptor status of SBC . The RR The present meta-analysis has several limitations. First, it is not possible to completely exclude the possibility that the HL itself carries with it an increased risk of second malignancy including SBC. Second, it is very difficult to quantify the possible effect of confounding factors such as lifestyle factors, personal risk, family history, etc. For example, Landgren et al. found increased RR (1.81) of breast cancer among HL patient with positive (vs. negative) family history of cancer . Third, We conclude that the current meta-analysis provided the most recent comprehensive estimate of the risk of SBC in a broad-range of HL survivors with inclusive analysis of relevant clinical and treatment variables. Based on the derived data where the median age at the diagnosis of SBC was 35.0 years and at a median latency of 17.7 years, screening recommendations for HL survivors need to be reemphasized. The results from the current meta-analysis support the favorable outcome of the risk-guided BC screening for such patients according to three prospective studies -76. It iAll authors declare that they have no competing interests.EMI, KMA, GAK, OAA, and MA Conception and design of the meta-analysis. EMI Study coordination and tasks’ assignment. KMA, and GAK Initial literature search. EMI, KMA, GAK, OAA, and MA Review of all potential studies. EMI, KMA, GAK, OAA, and MA Data extraction. EMI, KMA, GAK, OAA, and MA Assessing quality of included studies. EMI Statistical analysis. EMI, KMA, and GAK Investigating heterogeneity. EMI, KMA, GAK, OAA, and MA Preparation of the manuscript. EMI, KMA, GAK, OAA, and MA Reading the final manuscript. EMI, KMA, GAK, OAA, and MA Approval of the final manuscript. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2407/12/197/prepub


TP53 gene were shown to be associated with inflammation and inflammatory disease. The aim of this study was to investigate the interactions of miR-34b/c and TP53 Arg72-Pro polymorphisms on the risk of IA in a Chinese population. A total of 590 individuals (including 164 patients with IA and 426 controls) were involved in this study. The polymorphisms  were genotyped by polymerase chain reaction-restriction fragment length polymorphism assay and DNA sequencing. We found that the CC genotype of miR-34b/c rs4938723 was significantly associated with a decreased risk of IA compared with the TT genotype. Moreover, a significant gene interaction of the carriers with the combined genotypes of miR-34b/c rs4938723CC and TP53 Arg72Pro CG/CC/GG had a decreased risk of IA, compared with those carrying miR-34b/c rs4938723CT/TT+TP53 Arg72Pro GG/CG/CC combined genotypes. These findings suggest that the miR-34b/c rs4938723CC and TP53 Arg72-Pro polymorphisms may be involved in the susceptibility to IA.Several lines of evidence indicate that inflammatory processes play a key role in the happening and development of intracranial aneurysm (IA). Recently, polymorphisms in the  Intracranial aneurysm (IA) is a common disease with a high prevalence ranging from 1 to 5 percent in large autopsy studies –7. SAH rTP53 gene has an important function in cell cycle control, apoptosis, and maintenance of DNA integrity [TP53, located on chromosome 17p13 [TP53 gene is a frequent functional SNP that leads to a methionine proline conversion [TP53 Arg72Pro SNP results in a change in its protein structure reflected by its altered electrophoretic mobility [TP53 was reported to influence the p53-mediated inflammatory response [Both environmental and genetic factors are involved in the etiology of IA , 23. Sevntegrity –39. The ntegrity . TP53, lme 17p13 , 42 is 1me 17p13 . Codon 7nversion , 45. Themobility , and thiresponse .miR-34 family members, which compose 3 mature miRNAs that are encoded by 2 different pri-miRNAs [miR-34b/c, which may contribute to the susceptibility of HCC in a Chinese population [It is well known that p53 can regulate the expression of miRNAs, especially the i-miRNAs –54. The i-miRNAs . A potenpulation .miR-34b/c and TP53 Arg72-Pro were associated with the risk of IA. To test this hypothesis, we genotyped the 2 SNPs in a case-control study of 164 IA patients and 426 healthy controls in a Chinese population.In this study, we hypothesized that SNPs rs4938723 (T > C) in the promoter region of The study was performed with the approval of the hospital ethics committee, and written informed consent was obtained from all subjects participating in this study. The case-control study population contained 590 unrelated Chinese Han individuals including 164 patients ) with IA and 426 healthy controls ) living in Sichuan province of southwest China. Patients were recruited from the West China Hospital, Sichuan University from January 2008 to September 2009 who were newly diagnosed when they came for emergency because of SAH caused by the rupture of IA or just had general clinical symptoms such as headache or dizziness and diagnosed by DSA . The control group consisted of 426 healthy volunteers from a routine health survey in the same hospital during the same time as the patients. Subjects with any disease in nervous system or other serious illness were intentionally excluded. There was no significant difference between patients and control subjects in age distribution.μL EDTA-anticoagulated peripheral blood using a commercial extraction kit  according to the instruction manual. We used a polymerase chain reaction-restriction fragment length polymorphism assay to detect the genotype of the two SNPs . Primer sequences, reaction conditions, restriction enzymes  used, and length of resulting polymerase chain reaction products have been described previously [Genomic DNA was extracted from 200 eviously . Restrict test. Genotype and allele frequencies of miR-34b/c rs4938723 and TP53 Arg72Pro were obtained using Modified-Powerstates standard edition software. Hardy-Weinberg equilibrium was tested with chi-square test to compare the observed genotype frequencies among the subjects with the expected genotype frequencies. Genotype and allele frequencies of the two SNPs were compared between IA cases and controls using the chi-square tests. Odds ratios (ORs) and 95% confidence intervals (CIs) were used to assess the relative risk conferred by a particular allele, genotype, or the combined genotypes of miR-34b/c rs4938723 and TP53 Arg72Pro. Statistical significance was set at the P < 0.05 level. All the data were analyzed using the SPSS for windows software package version 13.0 .In this retrospective study, demographic and clinical data of both groups were compared by the chi-square test and miR-34b/c rs4938723 and TP53 Arg72Pro were successfully genotyped for 164 patients with IA and 426 controls. The clinical characteristics including sex, age of both groups, and the number of aneurysms in the cases enrolled in our study are shown in miR-34b/c rs4938723 and TP53 Arg72Pro are summarized in Tables miR-34b/c rs4938723 was significantly associated with a decreased risk of IA, compared with the TT genotype . However, no association between TP53 Arg72Pro and the risk of IA was observed for genotypic or allelic association analysis. We also examined the combined effects of miR-34b/c rs4938723 and TP53 Arg72Pro variants on IA risk. As shown in TP53 Arg72Pro CG/CC/GG had a 0.27-fold decreased risk of IA , compared with those carrying rs4938723CT/TT + TP53 Arg72Pro CG/CC/GG combined genotypes, while no other association between combined effects of miR-34b/c rs4938723 and TP53 Arg72Pro variants on IA risk was found. When stratification analysis was done by gender, no association was found (Tables The two SNPs of d Tables .miR-34b/c rs4938723 and TP53 Arg72Pro) and the risk of IA in a Chinese population. We found that the CC genotype of miR-34b/c rs4938723 was significantly associated with a decreased risk of IA, compared with the TT genotype. Moreover, a gene interaction of the carriers with the combined genotypes of miR-34b/c rs4938723CC and TP53 Arg72Pro CG/CC/GG had a decreased risk of IA, compared with those carrying miR-34b/c rs4938723 CT/TT + TP53 Arg72Pro CG/CC/GG combined genotypes. However, no difference was found when stratification analysis was done by gender for both miR-34b/c rs4938723CC and TP53 Arg72Pro.In this study, we investigated the association between two SNPs  synovial tissues [TP53 influences the p53-mediated inflammatory response. The role of p53 in innate immunity and the inflammatory response is now well established [in vivo and the alteration of miR-34b/c under an inflammatory microenvironment can be influenced by p53 [There are two ing site –54. Polying site . Severaling site –62. Whenp53 gene –65. Anot tissues –69. Fran tissues  reportedablished –72 and, ablished . Also a d by p53 .miRNA and p53 gene polymorphisms are associated with IA remain unclear, additional functional studies would provide valuable characterization of the molecular mechanisms by which miRNA and TP53  are involved in susceptibility to IA. However, our study provides evidence that miRNA and TP53 polymorphisms may play an important role in individuals' susceptibility to IA.Although the molecular mechanisms by which There were several limitations in our study. Firstly, detailed lifestyle that may be involved in the occurrence and development of IA was not available. Secondly, the number of subjects in the study was so small which might not be a good representative of the general population. Further studies, therefore, still need to be done.miR-34b/c rs4938723 was significantly associated with a decreased risk of IA, and a significant gene interaction of miR-34b/c rs4938723CT/TT and TP53 Arg72Pro CG/CC/GG was evident on the risk of IA. These findings suggest that miR-34b/c rs4938723 and TP53 Arg72Pro play a role in the formation, process, or rupture of IA. Nevertheless, our findings need to be replicated in larger, preferably multiethic population-based studies.In conclusion, we found that the CC genotype of