The ear has a reflexive property; therefore, various physical attributes may appear on the auricle when disorders of the internal organs or other parts of the body exist. Auricular diagnostics is an objective, painless, and noninvasive method that provides rapid access to information. Thus, the association between auricular signals and coronary heart disease (CHD) should be further investigated. A case control study was conducted to determine the predictive value of auricular signals on 100 cases of CHD  via visual inspection, electrical skin resistance measurement, and tenderness testing. The results showed that the presence of an ear lobe crease (ELC) was significantly associated with coronary heart disease. The “heart” zone of the CHD+ve group significantly exhibited higher conductivity on both ears than that of the controls. The CHD+ve group experienced significant tenderness in the “heart” region compared with those in the CHD−ve group in both acute and chronic conditions. Further studies that take into consideration the impact of age, race, and earlobe shape on ELC prevalence in a larger sample should be done. Yellow Emperor's Classics of Internal Medicine, which was published more than 2,000 years ago. The book states that the ear is related to all parts of the human body and internal organs and that all meridians converge at the ear. The Chinese Association of Acupuncture and Moxibustion was tasked by the World Health Organization to standardize acupoint locations to have a common language in the study and exchange of ideas. In May 1993, the “Chinese Standard Ear-Acupoint the Chinese Chart,” which consists of 91 acupoints, was established [The ear was first mentioned in the earliest Chinese medical book, The ear has a reflexive property; therefore, various physical attributes may appear on the auricle when body disorders exist. Such attributes include variations in shape, color, size, and sensation; appearance of papules, creases, and edema; and increased tenderness or decreased electrical conductivity , 3. The Qi, which occurs with aging [Other auricular signs associated with body functions also exist. For example, a published paper has reported that the presence of ELC has a high CHD predictive value . Meanwhith aging . From thth aging . Other pth aging . In addith aging .Despite the decline in age-standardized death rates of CHD patients in the last two decades, deaths due to CHD accounts for 70% of the mortality rate of heart disease . TherefoThis study aims to investigate the predictive value of auricular diagnosis on CHD. The hypotheses of this study are as follows: (1) visual inspection results indicated that patients with CHD exhibit a higher occurrence of ELC, appearance of edema ripples in the “Heart” region after pressing, and hair growth on the ears compared with community control group; (2) patients with CHD have lower electrical skin resistance on the “Heart” region of the ears than the community control group; (3) patients with CHD have significant tenderness on the “Heart” region of the ears than the community control group; (4) the degree of stenosis of the coronary arteries and the number of major epicardial arteries involved in CHD patients are associated with the presence of ELC; (5) a significant association exists between lipid profile (cholesterol and triglycerides) and the presence of hair growth or other auricular signals. The location of the “Heart” region in the Chinese Standard Ear-Acupoint Chart is displayed in n = 25), that is, newly diagnosed of having CHD within three months, whereas the rest were chronic CHD cases (n = 25). All CHD+ve cases were compared with the community subjects without CHD (CHD−ve group), who did not have any previous medical history of CHD or any experience of having cardiac-related symptoms including palpitation, chest pain, and/or dyspnoea. The control group was recruited from hospital staff, relatives, or friends of CHD patients. The participants in the case and control groups were matched by age and gender. Only the subjects who were aged 18 years or above were recruited, and patients with aural injury or infections were excluded from the study.This pilot study is a case-control study. Potential subjects were recruited from a regional hospital in Hong Kong (CHD+ve) and from the community (CHD−ve). Fifty subjects from each group were recruited through convenience sampling. The subjects from the CHD+ve group were recruited from the cardiac unit of the target hospital. Half of the 50 subjects suffered from acute episodes (n = 50) were compared with patients without evidence of CHD . Ear diagnosis was conducted in three ways: through inspection, palpation, and electrical detection. The observer was blinded to the grouping of the participants. Special auricular signals that were associated with coronary risks were observed and recorded on both ears.A simple health assessment was conducted , medical and family history, fasting cholesterol, triglycerides, and glucose) on the participants. Auricular signals that appear on patients with CHD , appearance of edema ripples in “Heart” region after pressing, presence of ELC, presence of ear hairs, and electrical resistance detected on the “Heart” region.The auricles were assessed based on the following procedures.Both auricles were observed for discoloration, appearance of edema ripples in “Heart” region after pressing, presence of ear hair, and ELC. ELC was graded according to the system modified from Patel et al. : grade 0An individual threshold was set for each subject before assessing the ear acupoints. The threshold was obtained by placing the acupoint detector on the “Shenmen” point and increasing the detection sensitivity until the sound, lights, or visual meter on the equipment indicates a high electrical conductance. The sensitivity was then slightly reduced until the “Shenmen” point was only barely picked up , 3. An eA pressure algometer (force gauge) with a unit range of 0 g to 500 g was used to apply force in the “Heart” zone using the “Shenmen” point as reference. The observed value (g) was recorded each time the subject starts feeling pain when the pointer of the instrument was applied on the acupoints during testing.A checklist was used to facilitate data collection. The data on coronary risk factors was collected. Increasing age, male gender, and heredity are the risk factors that cannot be changed; however, tobacco smoking, high blood cholesterol level, hypertension, obesity, and diabetes mellitus are modifiable risk factors.The findings of previous coronary angiography (within six months) for the CHD+ve group were retrieved to assess the severity of coronary artery stenosis (%) and the number of major epicardial arteries involved. The patients were divided into two groups: those with significant coronary atherosclerosis (≥50% stenosis in at least one of the major coronary arteries) and those with negligible or low degree stenosis (<50%), according to the criteria set by Kaukola et al. . The numThe inter-observer agreement of observations by the Principal Investigator and the Research Assistant (RA) was recorded for ten cases to assess the reliability of the auricular examinations. Digital photos of the auricles were obtained to examine the inter-observer agreement of the ELC grading using Kappa's statistic, thereby increasing the validity of the findings . Both tht-test was used to compare the mean scores of the continuous variables between the case and control groups, as well as between CHD patients with acute episodes and those with chronic conditions, when appropriate. These variables were namely the body mass index, lipid profile, ELC%, and electrical resistance readings of individual acupoints. A multivariate logistic regression analysis was conducted to examine the association of the cardiovascular status of the subjects with the auricular signals after controlling for potential confounders. The auricular signals included the presence or absence of ELC, hair growth, edema ripples, tenderness, and the conductivity of the “Heart” region. Analyses were conducted using SPSS Statistics 19. A value of P < 0.05 was considered statistically significant.Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) with 95% confidence intervals (Cls), as well as odds ratios of auricular signals  that predict the CHD were computed. ELC grading was converted to percentage (from 0% to 100%), and a receiver operator characteristics (ROC) curve was plotted to produce a table of sensitivity and specificity for different cutoff scores. Kappa's statistic was computed to determine agreements on CHD status (+ve versus −ve) and those classified by different cutoff points of ELC%. Chi-square test was used to determine the association between the auricular signals and CHD (+ve versus −ve) and between auricular signals and the degree of coronary stenosis . The Student's Ethical approval from the hospital and the university was sought. Written informed consent was obtained from every eligible person who agreed to participate. The purpose and procedures of the study were explained verbally and in writing to the participants. Participation in the study was on a voluntary basis, and all participants were assured that they have the right to refuse or withdraw from the study at any time. Personal information and data remained confidential and anonymous.P > 0.05) with slightly lower mean arterial pressure  than those in the CHD-ve group. Therefore, these two variables were adjusted for the subsequent multivariate analyses  and 50 from the community (CHD−ve as controls). The participants in the case and control groups were matched by age and gender. The mean age of the participants was 65.32 years (S.D. = 14.12), with 54 males and 46 females. The mean ages of the participants for the acute and chronic cases were 64.92 years (S.D. = 14.55) and 65.56 years (S.D. = 14.76), respectively, with no significant gender distribution between groups. No significant differences were observed in the BMI, presence of comorbid illness, and the family history of myocardial infarction between the groups. However, participants in the CHD+ve group have more ex-smokers/smokers  were compared with those of patients without evidence of CHD (CHD−ve). P < 0.05; P < 0.05), whereas 43.1% versus 24.0% was noted on the right ear (P < 0.05) in the CHD+ve and CHD−ve groups, respectively. In addition, more participants in the CHD+ve group exhibited hair growth on ears than those in the CHD−ve group, especially on the right ear (P < 0.01). The results from the stratified analyses indicated that the average percentage of ELC across the ear lobe among cases with acute condition was significantly different from the control group, whereas the chronic cases were found to have significant hair growth on right ear.The presence of ELC, which is the primary auricular signal for CHD prediction in this study, was significantly associated with CHD. More participants in the CHD+ve group showed the presence of ELC compared with those in the CHD−ve group . Bilateral ELCs were present in 45% (n = 45) of the participants, whereas 13% showed one-sided ELC. The ELC sign on the right  and left ears  was associated with increasing age, but was independently associated with gender. Edema ripples in the “Heart” region after pressing appeared in most participants of the CHD+ve group (P < 0.05). Among these CHD+ve cases, the most commonly involved artery with blockage was the left anterior descending , followed by the right coronary artery , left circumflex , posterior descending artery , and posterior lateral branch . Among these arteries, the degree of RCA stenosis was significantly associated with the presence of ELC on either the right (P < 0.001) or left ear (P < 0.05). Moreover, the number of major epicardial arteries involved was also associated with the presence of ELC  and edema around the “Heart” zone (P < 0.05) on the right ear.A significant relationship between the presence of ELC on the right and left ears was observed in all cases  or left ear .Among these auricular signs, ELC sensitivity was 61.0%, specificity was 58.0%, and PPV was 59.3%. An ROC curve was generated to produce the best cut-point analysis. The use of 35% ELC across the ear lobe was the most desirable cutoff value for predicting CHD status, which was supported by a fair agreement of Kappa's statistic in either the right . The results from the stratified analyses revealed a significant change in the conductivity of the “Heart” zone in chronic CHD+ve cases (P < 0.01), but not in acute cases. The conductivity of the “Heart” zone exhibited a sensitivity of 49.0%, a specificity of 80.0%, and a PPV of 71.7%. However, no association was observed between the electrical conductivity of the “Heart zone” and the degree of stenosis of the coronary arteries and the number of major epicardial arteries involved.The “Heart” zone of the CHD+ve cases had a significantly higher conductivity on both ears  compared with the control group (P < 0.001) compared with those in the CHD−ve group. A significant relationship was noted between the average tenderness (%) on both ears . The results from the stratified analyses indicated that these two signals were found significant in cases with either acute (P < 0.01) or chronic condition (P < 0.01). The result from the tenderness testing of the “Heart” zone showed a sensitivity of 92.0%, a specificity 75.0%, and a PPV of 78.6%. No association was observed between tenderness of the “Heart” zone and the degree of stenosis of the coronary arteries and the number of major epicardial arteries involved.The participants in the CHD+ve group experienced significant tenderness in the “Heart” region in both ears (P < 0.05) were retained in the final model (P < 0.001) and correctly classified in 88.0% of the subjects. The result of the Hosmer and Lemeshow test indicated that the model effectively fits with the data (P = 0.081). In this final model, each 1% increase in the ELC occurrence on the left side of the ear will increase the risk of CHD by 1.017 times . The presence of tenderness  and conductivity  in the “Heart” region of the left ear was likewise significantly associated with the CHD status after the adjustment of the smoking status and MAP readings. However, when the relationships among the three auricular signals were examined, all their associations on the left and right ears as well as in the acute and chronic cases were not significant.Multivariate logistic regression analysis was conducted to examine the association of the cardiovascular status of the subjects with a number of auricular signals after controlling for the significant risk factors (smoking status and MAP). Age and gender were not included in the model because the cases and controls were matched according to these attributes. Backward logistic regression was used, and auricular signals that demonstrated statistical significance  than the control group. The electrical resistance in the corresponding auricular points decreases when a disease or disorder is present in the body, and areas with low electrical resistance than that of the standard are considered either positively or highly conductive electrical resistance points . SeveralThe participants in the CHD+ve group experienced significant tenderness in the “Heart” region than those in the CHD−ve group in both acute and chronic cases as detected by the pressure algometer (force gauge). The degree of tenderness is usually related to the severity of the condition; the more sensitive the point, the more severe the disorder , 40. In Based on the multivariate logistic regression analyses, the presence of ELC on the left side of the ear as well as the tenderness and conductivity in the “Heart” region of the left ear were notably associated with the CHD status after the adjustment of the smoking status and MAP readings. The occurrence rate of ELC on the left side of the ear was found to be slightly higher than that on the right ear. This phenomenon may possibly be explained by the reflex pathway of the “Heart”, which is more likely to occur on the left side of the ear. This trend is similar to the pictograph of foot reflexology, which shows that the “Heart” is located on the left foot and not on the right foot. Moreover, the said phenomenon may be associated with the fact that the LAD artery in these subjects was most commonly involved in blockage (75.0%).The results of this study can advance our knowledge in understanding the association between specific auricular reflective signs and CHD status. Such information will be a valuable background data to support future studies for screening vulnerable populations with CHD risk, such as those with family history, hypertension, hyperlipidemia, sedentary lifestyle, and obesity. Auricular diagnosis has a pre-diagnostic value and is important in the secondary level of prevention. Prognosis may be improved by early preventive measures if CHD status can be identified at an earlier stage. The examination of the auricles may be integrated into the routine clinical examination of a patient to increase the predictive accuracy of the coronary risk profile for earlier introduction of preventive measures, such as control of hypertension and smoking, before the disease advances. This process may help detect CHD in its early stages and shed some light on the early development of coronary atherosclerosis. ELC may be a useful indicator for the presence of CHD in the preoperative assessment of patients, particularly in emergency situation, where little or no history and investigations are available. The use of ELC may improve the clinician's ability to estimate the probability of CHD in patients before they undergo diagnostic testing and to predict the operative risk , 20. GivAlthough the association between ELC and CHD is suggested in this study, the mechanisms leading to the concurrent development of ELC and CHD and the time of onset remains uncertain. A prospective cohort study could be conducted in the future to follow-up the newly diagnosed CHD+ve cases and to identify changes in the auricular signals that may have appeared during disease progression. This study was conducted on a relatively small sample. Therefore, further investigations must be performed with a larger sample to validate the use of auricular signs as predictors that assist in the diagnosis of CHD. Due to resource implications, the CHD−ve status can only be verified by self-report of the participants in the control group who do not have any previous medical history of CHD or any experience of having cardiac-related symptoms, for example, palpitation, chest pain, and dyspnoea. The prevalence of ELC may be governed by ethnical differences; thus, more work is required before these results can be extrapolated to other ethnic groups.The predictive value of auricular signals on CHD, which was detected by visual inspection, electrical skin resistance measurement, and tenderness testing, was observed in the Chinese population in this study. Such findings can advance our knowledge on the integrated approach that combines Chinese and Western models of care for diagnosing patients with underlying CHD. Further studies should be conducted to ascertain the diagnostic value of AT on CHD.


NAT1) genotype, a drug-metabolizing enzyme coding gene, and bladder cancer in Lebanese men. A case-control study  was conducted in two major hospitals in Beirut. Cases were randomly selected from patients diagnosed in the period of 2002–2008. Controls were conveniently identified and selected from the same settings. Data was collected using interview questionnaire and blood analysis. NAT1 genotypes were determined by PCR-RFLP. Statistical analysis revolved around univariate, bivariate, and multivariate logistic regression models, along with checks for effect modification. Results showed NAT1∗14A allele, smoking, occupational exposure to combustion fumes, and prostate-related symptoms, to be risk factors for bladder cancer. The odds of carrying at least one NAT1∗14A allele are 7 times higher in cases compared to controls . A gene-environment interaction was identified for NAT1∗14A allele with occupational exposure to combustion fumes. Among carriers of NAT1∗14A allele, the odds of bladder cancer dropped to 2.03 from 3.72. Our study suggests NAT1∗14A allele as a possible biomarker for bladder cancer. Further research is recommended to confirm this association.In Lebanon, bladder cancer is the second most incident cancer among men. This study investigates a possible association between N-acetyltransferase 1 ( Globally, the prevalence of bladder cancer ranks nine and accounts for 3.3% of all malignancies . As a geGeographical and ethnic differences in incidence may be attributed to differences in environmental and genetic factors. Historically, the majority of genetic studies investigating bladder cancer focused on the nuclear genome such as oncogenes, tumor suppressor genes, and subsequent cellular events such as DNA repair, genomic instability, disturbance of gene regulation, alterations of signal transduction, and apoptotic pathways , 8. In tNAT1) gained attention when, contrary to NAT2, was shown to be expressed in a wider number of tissues including bladder epithelia [NAT1 endogenous substances are not well known although there is some evidence that suggests a role for the enzyme in folate metabolism [NAT1 was originally described as monomorphic based on its selectivity for p-amino-salicylic acid and metabolic yield [Of particular importance are N-acetyltransferase Phase II enzymes known to mediate N-acetylation and O-acetylation reactions of arylamine derivatives. N-acetyltransferase 1  [NAT1 genetic polymorphism and bladder cancer among Lebanese men residing in Lebanon.The current research was triggered in view of the alarmingly high incidence of bladder cancer in Lebanon, and based on an unusually high  allele) , 23, 24.A case-control study was conducted in two major medical centers in the Capital Beirut: St. Georges Hospital and Bahman Hospital. 54 cases and 106 hospital controls were selected. Originally, a total of 284 bladder cancer patients were identified from medical records of both hospitals between 2002 and 2008. Contact information was obtained from the archives of both hospitals. Excluded patients were females, men under age of 50, deceased patients before the beginning of the study, patients with missing contact information, in addition to patients who refused to participate. The remaining patients were included in the study as detailed in NAT1*14A among cases of 21.4%, based on data observed in earlier studies [The final sample size was guided by power analysis that revolved around the following: power of 80%, type 1 error of 5%, estimated OR of 3, and a proportion of exposure of  studies , 23, 24.Cases were Lebanese males above the age of 50 with a histologically confirmed bladder cancer diagnosed at Saint Georges University Medical Center, and Bahman Hospital between 2002 and 2008. Cases were randomly selected as per year of reporting from the most recent. Controls were conveniently selected from the same settings. These were subjects attending to both settings either for a social visit or a routine checkup. They were Lebanese males, 50 years or older, with no present or previous history of cancer, or any other systemic illness or chronic disease.The study excluded women, non-Lebanese, and all subjects that are less than 50 years of age for both cases and control groups. In addition, all first-degree relatives were excluded from both cases and control groups. In the control group, all subjects with history of bladder cancer, or other types of cancer or urinary infections, or chronic diseases, were excluded.P values, and confidence intervals were computed at a type I error alpha value of 5%. The final model incorporated the exposure and confounding variables that displayed the most significant odds ratios.University of Balamand and Hospitals IRB approvals were obtained prior to conducting the study. Data collection involved combining structured face-to-face interview questionnaires and blood withdrawal. The questionnaire gathered information on the following: age, family history, smoking habits, alcohol consumption, dietary habits, chronic diseases and urinary infections, use of hair dyes, and occupation. The questionnaire was administered by a trained research assistant in native Arabic. After collection, data were coded, entered, and analyzed using the Statistical Package for Social Sciences . Univariate analysis consisted of frequency and percentage distributions for the different categorical variables in the study. Means, SDs, and ranges were computed for the different continuous variables, with checking for normality and outliers. Bivariate analysis mainly used Chi-squared and Fisher's exact test to check for an association between the main outcome variable (urinary bladder) and various exposure and confounding variables. The purpose of this analysis was to examine crude associations and to check for potential confounders and effect modifiers. Multivariate analysis consisted of a backward multivariate logistic regression model, in which analysis included the different exposure and confounding variables that yielded significant results during bivariate analysis. Odds ratios, 2 mL of blood was collected in EDTA-treated tubes from each bladder cancer patient and control subject and stored at 4°C until DNA was extracted. To each blood sample 1 mL of 100% Ficoll was added to improve isolation of buffy coats containing the nucleated white blood cells. This coat appeared after centrifugation at 2500 g for 10 minutes. DNA extraction was performed using a ready-to-use extraction kit (Qiagen) according to manufacturer's instructions and stored at −20°C until genotype analyses.NAT1 variant alleles as described [NAT1 was amplified by PCR in a 30 μL reaction using 150 ng of genomic DNA as a template, 10 mM Tris-HCL, pH 8.3, 50 mM KCL, 1.5 mM MgCl2, 0.2 dNTP, 100 ng of primers NAT1-f (−10 to 29) 5′-TTA GGA ATT CAT GGA CAT TGA AGC ATA TCT TGA AAG AAT-3′ and NAT1-r (1148 to 1127) 5′-GCT TTC TAG CAT AAA TCA CCA A-3′, and 0.75 unit of Taq DNA polymerase (Fermentas). The mixture was subjected to a 5 min denaturing at 94°C, followed by 30 cycles of 1 min at 94°C, 1 min at 55°C, 1 min at 72°C, and concluded with a 7 min final extension at 72°C. Following PCR, an aliquot of the product was digested with AatII to confirm amplification. Nucleotide substitution G350, 351C (identifier of NAT1*5) was detected by digesting NAT1 PCR product with 10 units of BanI as described above for AatII. Digested samples were run on a 2% agarose gel containing ethidium bromide and examined under UV light.PCR-RFLP assays were used to characterize wild-type and escribed . Briefly1095A, C559T, G560C, and T1088A, nested PCR was performed. Briefly, a 0.5 μL sample of amplified NAT1 was used as template in a 20 μL reaction containing 10 mM Tris-HCL, pH 8.3, 50 mM KCl, 1.5 mM MgCl2, 0.2 mM each dNTP, 200–350 ng of appropriate primers, and 0.75 units Taq DNA polymerase. The mixture was amplified according to conditions shown in μL of C1095A nested PCR product was digested overnight at 37°C with 2 units of MseI and then was run on a 5% Metaphor gel. All other digested samples were run on a 2% agarose gel containing ethidium bromide and examined under UV light  years compared to 65.6 (±11.3) years in controls. Most subjects had completed junior-high school (around 8 years of education). The average monthly income in both groups was around US $1,800. The majority of cases and controls resided in Beirut (33.3% versus 79%), and Mount Lebanon (46.5% versus 15%) . Most ofNAT1*14A , NAT1*14B (13% versus 4.7%), and NAT1*15 (48.1% versus 40.6%). Moreover, cases were less likely than controls to have the clustering of the following alleles: NAT1*10 , NAT1*3 (13 versus 13.2%), and NAT1*4   . NAT1*5 P values of 0.1 or lower, both with and without testing for potential gene-environment interaction. The odds of having an income > 1000 US$/month were 7.45 times higher in cases compared to controls, while the odds of an income ranging between 500 and 1000 US$/month were 2.14 times higher in cases than controls. Smoking, prostate-related symptoms, and NAT1*14A allele were found to be significantly independent risk factors for bladder cancer. Occupational exposure to combustion fumes was also found to be an important risk factor, but not statistically significant. When comparing odds ratios in cases versus controls, the odds of NAT1*14A allele clustering were 6.94 times higher in cases, while the odds of NAT1*10 were 0.74 times (lower in cases compared to controls). In addition, the odds of smoking were found to be 1.02 times higher in cases; the odds of exposure to occupational vapors and fumes were 4.34 times higher in cases; the odds of prostate-related symptoms were 7.81 times higher in cases, compared to controls  and to have a significantly higher clustering among cases compared to controls . We also found that the odds of NAT1*10 allele carriers are lower among cases compared to controls. In addition, our results present smoking and prostate-related symptoms as independent predisposing factors for bladder cancer.Identification of the underlying causes of bladder cancer in Lebanese men is becoming an urgent need , 27–29. NAT1 affecting bladder cancer risk is not particularly new. Our study, however, reenforces this hypothesis by further investigating the association in a population with both a remarkably high bladder cancer incidence and a reported unusual high frequency of a suspected allele [NAT1*14A frequency in Lebanese is considered significantly high when compared to other ethnic groups in the literature. For example, NAT1*14A frequency was reported to be 2.8% in a predominantly Caucasian population [NAT1*14A with higher risk, and NAT1*10 with lower risk, are consistent with a previously conducted case-control study among a selected German population [The hypothesis of d allele . Our find allele . The reppulation , 1.3% ampulation , 1.7% inpulation , and 3% pulation . Our finpulation . NAT1*14A individuals have been found to date, including the present study, NAT1*14A has been reported to produce a slow acetylation phenotype based on studies in recombinant enzyme expressed in E. coli and blood mononuclear cell lysate [NAT2 slow acetylator phenotype is reported to be more prominent [NAT1 is the main NAT expressed in bladder epithelia and cuboidal epithelium of proximal convoluted tubules in the kidney. Recent studies have suggested a novel role for this enzyme in cancer cell growth through folate homeostasis. It is upregulated in several cancer types, and overexpression can lead to increased survival [NAT1 and cancer cell growth remain to be explained. Potentially, in a population with higher NAT1*14A frequency such as this one, NAT1 slow acetylation may compete poorly with Phase I bioactivation pathways for arylamines and hence contribute to a higher bladder cancer risk. For NAT1*10, on the other hand, reports remain inconsistent. While some studies have reported no change in enzymatic activity for NAT1*10, other reports suggest its association with a higher acetylation activity in colon, bladder, and liver tissues [NAT1*10, the low power of the interaction analyses precludes us from making conclusions. Phenotypes may vary with organ/tissue and may be dependent upon other endogenous and environmental factors. Evidence exists for heterogeneity within the “slow” acetylator phenotype [Although no homozygous l lysate , 23. Rodl lysate , 34. Whirominent , NAT1 issurvival , 37. Man tissues , 23, 38.henotype , 40. NAT1*14A had a higher bladder cancer risk compared to noncarriers. This is an important finding that needs followup using quantitative more accurate measurement tools for prostate-related symptoms assessment. Potentially, partial blockage of bladder outflow, due to a prostate disorder, may increase contact time of chemicals with mucosal cell lining, thereby increasing odds of malignant transformation, particularly in carriers of the slow NAT1*14A phenotype. On the other hand, our analysis suggests a lower risk for NAT1*14A carriers exposed to occupational combustion fumes compared to noncarriers. This needs further investigation before drawing conclusions. It may relate, in part, to differences in chemical exposure, other genetic factors related to metabolism, or potentially the small sample size. Our study did not investigate schistosomiasis as a potential risk factor since this infection is very rare in Lebanon and may be more relevant in countries like Egypt [ When we tested for interactions, smoking remained to be a significantly independent risk factor. Patients with prostate-related symptoms carrying ke Egypt . Schistoke Egypt . The majNAT2, that may modify bladder cancer risk, in addition to other risk factors not investigated by the current study. This is mainly due to limitations in both sample size and budget. Further studies should be conducted in the future to address these points. An important limitation worth noting is related to a possible misclassification of the NAT1*4-wild type allele. NAT1*4 may include other mutations not tested for in the current study. This may bias our risk estimates toward the null. However, many of these nucleotide substitutions and corresponding NAT1 alleles are reported to be very rare and reflect nucleotide diversity (frequency of 0.003–0.005) rather than genetic polymorphism, with many of them silent [Several limitations could have affected the results of the current study. The external validity of this case-control study is limited due to the selection of a hospital-based sample that might overshadow the ability to generalize to the overall population. A larger sample size is also needed for increasing the power of this study to be able to demonstrate a higher confidence when interpreting the observed results of effect modifiers. Although our sample size was guided by power analysis, it would have been more optimal to include larger number of cases . This was not possible due to the high proportion of refusals, deceased, and subjects with no contact information . Another limitation of the study is the possibility of other genetic polymorphisms, such as m silent . At the On the other hand, several strengths should also be highlighted in this case-control study. The quality of the measures used is high, particularly due to combining two formats of data collection: interview questionnaire and genetic predisposition analysis. No major challenges existed for adjusting for missing and non-response data. In addition, several multivariate models were analyzed before determining the best-fit models used. All the different constructed models yielded similar patterns in the observed results, which reflect the high internal validity of the observed findings reported herein. NAT1*14A influences bladder cancer risk in Lebanese males. Although the observed effects may be moderate, the suggested association between incidence and genetic variation may account for a substantial proportion of cases in the Lebanese context. On one hand, the NAT1 enzyme was recently reported as a biomarker of cancer, and its overexpression was found to be associated with increased resistance to chemotherapy [NAT1 14A may be a viable candidate for preclinical and clinical evaluation, as well as for drug development. On the other hand, early screening and detection of NAT1*14 in high-risk groups, such as smokers, may be important for Lebanese in view of the double burden of the two highest cancer incidents: lung and bladder. We recommend building upon these observations by carrying out larger multicenter studies incorporating larger sample sizes. Although individual risks associated with N-acetylation genotypes are small, they do increase when considered in conjunction with other metabolizing-enzyme genes, susceptibility genes, and chemical exposures. Because of the relatively high frequency of NAT1*14A in the Lebanese population, the attributable risk of bladder cancer in this population may be high. However, future studies must be directed toward investigating the contribution of the combined effect of other drug-metabolizing enzymes, particularly NAT2 acetylator polymorphism. In conclusion, this is the first case-control study in Lebanon attempting to explain the high incidence of bladder cancer. Our results suggest that otherapy . This fi


Vitamin D receptor (VDR) polymorphisms with urolithiasis risk in different ethnic groups. However, the results were inconclusive. To evaluate a more precise estimation of the relationship, a meta-analysis was performed.Several studies analyzed the associations of Pubmed, EMBASE, Wanfang Database, China National Knowledge Infrastructure (CNKI) and Weipu Database were searched. Data were extracted independently by two investigators. Odds ratios (ORs) with 95% confidence intervals (CIs) were used to assess the strength of associations.ApaI, BsmI, FokI, and TaqI polymorphisms and urolithiasis risk were observed. However, sensitivity analyses for BsmI and FokI polymorphisms indicated that the results were not reliable and credible. In addition, there was a significant association of the ApaI-TaqI haplotype with urolithiasis risk.Twenty-three case–control studies were included in this meta-analysis. Significant associations between ApaI and TaqI polymorphisms in VDR gene were associated with urolithiasis risk.This meta-analysis suggested that  Urolithiasis is one of the most prevalent uronephrologic disorders and affects approximately 10% of individuals in western countries . PreviouRecently, Elkoushy and coworkers found that patients with urolithiasis had a high prevalence of inadequate vitamin D . ExpressVDR gene is located on chromosome 12q12-14. Four single nucleotide polymorphisms (SNPs) of the VDR gene have been widely studied [ApaI, BsmI, and TaqI are located between the 8 and 9 exons in the 3′-untranslated region (UTR), and shown to be in strong linkage disequilibrium (LD) [FokI, which is located at the translation starting codon. Many studies investigated the associations between these polymorphisms of VDR gene and the risk of urolithiasis [VDR gene variants and urolithiasis risk.The human  studied . ApaI, Bithiasis -32. HoweWe performed a systematic search of Pubmed, EMBASE, Wanfang Database, China National Knowledge Infrastructure (CNKI) and Weipu Database to find relevant studies. The search terms were used as follows: (urolithiasis or kidney stone or kidney stone disease) and (Vitamin D receptor or VDR) and (polymorphism or mutation or variant). Last search was updated in October, 2012. No language restriction was imposed. The reference lists of searched articles and relevant reviews were all perused to find additional eligible studies.ApaI, BsmI, TaqI, and FokI polymorphisms in VDR gene and urolithiasis risk, (2) using a case–control design, and (3) genotype distributions in both cases and controls should be available for estimating an odds ratio (OR) and 95% confidence interval (CI).Two reviewers (Zhang and Nie) independently screened titles and abstracts of all studies for relevancy. Disagreements were resolved by discussion. Studies included in this meta-analysis based on the following selection criteria: (1) evaluation of the VDR polymorphisms were not analyzed or the outcome was not urolithiasis risk, (2) not case–control studies, such as the design based on family or sibling pairs, (3) not reported genotype frequencies or number, (4) abstracts or reviews, and (5) non-clinical study. For overlapping studies, the one with the largest sample size was included.Studies were excluded if one of the following existed: (1) the VDR gene, genotyping method, and genotype number in cases and controls. We contacted the corresponding authors if more data was needed.The full manuscripts of eligible studies were reviewed by two investigators (Zhang and Nie) independently. Any discrepancy was resolved by discussion or a third author (Jiang) would assess the articles. The following information was collected from each study: the first author’s name, year of publication, original country, ethnicity, age group, hypercalciuria in the urolithiasis group, composition of stone, sample size, the polymorphisms in The quality of the studies was assessed by two investigators (Zhang and Nie) independently. The predetermined criteria were modified from a previous review . These sApaI, BsmI, FokI, and TaqI polymorphisms and urolithiasis risk was measured by ORs and 95% CIs. OR1, OR2, and OR3 were calculated for the genotypes: 1) AA vs. aa (OR1), aA vs. aa (OR2), and AA vs. aA (OR3) for the ApaI, 2) bb vs. BB (OR1), bB vs. BB (OR2), and bb vs. bB (OR3) for the BsmI, 3) ff vs. FF (OR1), fF vs. FF (OR2), and ff vs. fF (OR3) for the FokI, and 4) tt vs. TT (OR1), tT vs. TT (OR2), and tt vs. tT (OR3) for the TaqI, respectively. The statistical significance of OR was analyzed by Z test. These pairwise differences were used to indicate the most appropriate genetic model as follows: if OR1 = OR3 ≠ 1 and OR2 = 1, then a recessive model was suggested; if OR1 = OR2 ≠ 1 and OR3 = 1, then a dominant model was suggested; if OR2 = 1/OR3 ≠ 1 and OR1 = 1, then a complete overdominant model was suggested; if OR1 > OR2 > 1 and OR1 > OR3 > 1 (or OR1 < OR2 < 1 and OR1 < OR3 < 1), then a codominant model was suggested [When the data from at least three similar studies were available, meta-analysis was performed. The strength of the associations between the uggested ,35. OnceQ-test and I2 test. I2 values of 25%, 50%, and 75% were nominally assigned as low, moderate, and high estimates. Departure from Hardy-Weinberg equilibrium (HWE) in controls was tested by the Chi-square test. Subgroup analyses were conducted by ethnicity, calciuria level, and age group. Sensitivity analyses were performed by excluding the studies not in HWE and the studies with low quality, respectively. Funnel plot was used to assess potential publication bias. Publication bias was also investigated statistically via Egger’s test [The between-study heterogeneity was assessed by the Chi square-test based Cochrane r’s test .P value < 0.05 was considered statistically significant, except for test of heterogeneity where a level of 0.10 was used.All statistical tests were performed by using STATA 11.0 software . A VDR polymorphisms. After reading the full texts of the remaining 32 articles, 9 articles were excluded due to irrelevant to urolithiasis risk, no useful data, and reduplicate study. Finally, a total of 23 case–control studies met our inclusion criteria. There were 11 studies on ApaI, 8 on BsmI, 14 on FokI, 13 studies on TaqI. Four studies reported the haplotype of ApaI and TaqI polymorphisms. There were 14 studies of Asians and 9 studies of Caucasians. Seventeen studies were performed in adults, 3 in children, and 1 did not offer detailed information. Three studies only included patients with hypercalciuria, 6 studies included hypercalciuria patients partly and the data for these patients could be extracted, and 14 studies did not report detailed information. Quality scores for the each study ranged from 3 to 6. The characteristics of each study are presented in Table Figure ApaI polymorphism and urolithiasis risk. Total sample sizes in urolithiasis and control groups were 1584 and 1853. The estimated OR1, OR2 and OR3 were 1.47, 1.30, and 1.04, respectively (Table P = 0.002). In the subgroup analysis by ethnicity, a significant association was found among Asians  but not among Caucasians . Subgroup analysis was also performed according to the calciuria level. However, no significant increased risk of urolithiasis was found among hypercalciuric patients  (Table P = 0.02). Sensitivity analysis was performed by excluding the studies that did not show HWE. The result was statistically significant . Sensitivity analysis was also performed by excluding the low quality studies. The result was similar . The shape of the funnel plot showed symmetric .Eleven studies determined the association between VDR BsmI polymorphism and urolithiasis risk were included in this meta-analysis. The estimated OR1, OR2 and OR3 were 1.79, 1.85, and 0.96, respectively (Table P = 0.04)  but not among Asians  (Table P = 0.003). In the stratified analysis by ethnicity, no significant association was found among adults . However, sensitivity analyses conducted by excluding the studies not in HWE or low quality studies did not find the significant association between VDR BsmI polymorphism and urolithiasis risk (Table P = 0.461).Eight studies (1210 cases and 1160 controls) that identified the association between VDR FokI polymorphism, fourteen studies including 2266 cases and 2418 controls were included in this meta-analysis. OR1, OR2, and OR3 were 1.59, 1.38, and 1.04, respectively. These estimates suggested a dominant genetic model. Therefore, the original grouping was collapsed, and ff and fF were combined, in accordance with a dominant model, into a f carrier group, the latter of which was compared with the FF genotype group. The pooled OR was 1.48    (Table P = 0.009). Sensitivity analyses found that the significant result was altered when the low quality studies or studies with Hardy-Weinberg disequilibrium were omitted (Table P = 0.081).For the ed Table . Egger’sVDR TaqI polymorphism and urolithiasis. The estimated OR1, OR2 and OR3 were 1.24, 1.33, and 0.92, respectively. These pooling estimates suggested a dominant genetic model. The pooled OR in the dominant genetic model was 1.30   but not Caucasians . Furthermore, patients with hypercalciuria had a significant increase of the risk of developing urolithiasis . In addition, significantly increased urolithiasis risks were found among adults . Sensitivity analysis did not change the result (Table P = 0.122).Thirteen studies including 1744 patients and 1944 controls addressed the association between VDR ApaI-TaqI haplotype in urolithiasis risk was analyzed. The sample sizes in case group and control group were 830 and 1207, respectively. Carling et al. [P = 0.007). In the subgroup analysis by ethnicity, a significant association was found among Asians . In the stratified analysis of age group, we also found a significant association . Symmetrical funnel plot was obtained .In 4 out of 23 studies, the role of g et al.  reportedd Figure . PublicaVDR polymorphisms and urolithiasis risk. We found that ApaI polymorphism was a risk factor for developing urolithiasis. This result indicated that the carriers of the AA or Aa genotype had a 34% increased urolithiasis risk compared to those individuals with the aa genotype. Sensitivity analysis did not change this result, suggesting the solidity of our result. In the stratified analysis by ethnicity, the significant association was found in Asian population but not in Caucasian population. In addition, BsmI and FokI polymorphisms were also significantly associated with urolithiasis risk in the overall population. However, results from sensitivity analyses suggested that these results were not statistically robust. Therefore, these results should be interpreted with caution and more studies are needed to evaluate the effect of BsmI and FokI polymorphisms on urolithiasis risk. Specifically, we noted that hypercalciuric patients who harbored b allele may have increased urolithiasis risk. As for the TaqI polymorphism, a significant association was observed between this polymorphism and the risk of urolithiasis. Sensitivity analyses further strengthened the validity of this result. When subgroup analysis was conducted according to the ethnicity, the significant association was showed in Asians and lack of a significant association was detected in Caucasian population. In the hypercalciuria subgroup analysis, a 56% increased risk of urolithiasis was found in patients with hypercalciuria. Collectively, these results suggested that VDR BsmI and TaqI polymorphisms may play unique roles in the etiology of hypercalciuric urolithiasis. However, because the number of studies included in the hypercalciuria subgroup analyses was small, the results lacked sufficient reliability to confirm these associations in a definitive manner. Thus, future studies with larger sample sizes are needed to confirm our results. Furthermore, we assessed the association between the haplotypes of the ApaI and TaqI polymorphisms and urolithiasis risk. The result from this meta-analysis suggested that At haplotype may play a role in urolithiasis susceptibility, especially in Asians. Taken together, our results exhibited significant associations between ApaI and TaqI polymorphisms and urolithiasis risk in Asian population but not in Caucasian population. There are at least four reasons to explain the ethnic difference. First, the number of Caucasian patients was small. There were only 384 and 644 Caucasian patients for these two polymorphisms, respectively. It was therefore possible that the observed ethnic difference was due to chance. More studies with Caucasian population are required to validate the effect of ethnic differences. Second, higher heterogeneity was observed in the Caucasians subgroup (I2 = 40%) but not in the Asians subgroup (I2 = 0%) in TaqI polymorphism. This may distort the result. Third, urolithiasis is a complex disease. Both genetic and environmental factors could affect the risk of urolithiasis in different populations. It is possible that different urolithiasis risks in Asians and Caucasians were due to exposure to various environmental factors. To data, there was no reported study which was performed to assess the effect of VDR-environment interactions on urolithiasis risk in different ethnicities and regions. Therefore, epidemiologic studies should be designed to examine these associations in the future. Fourth, ethnic difference in the VDR gene allele frequencies may also result in this difference. For example, the A allele of the ApaI, occured with lower frequency in Caucasians when compared to Asians, while the t allele of the TaqI had a higher frequency in Asians compared to Caucasians [BsmI polymorphism. These results indicated that these polymorphisms were significantly associated with increased urolithiasis risk in adults. We did not evaluate the associations between VDR polymorphisms and urolithiasis risk in children due to insufficient data. More studies using children population are needed to determine the associations between VDR polymorphisms and urolithiasis risk.This present meta-analysis systematic investigated the associations between VDR, Xi and colleagues silenced the VDR gene in the kidneys of GHS rats [VDR knockdown in the kidney can upregulate the expression of transient receptor potential vanilloid receptor subtype 5 (TRPV5) in GHS rats [VDR was one of the most studied candidate genes for urolithiasis. Carling et al. [VDR gene for the BsmI polymorphism may be associated with higher urinary calcium excretion. Hypercalciuria was a risk factor of urolithiasis. It was thus possible that b allele carriers may have higher urolithiasis risk than B allele carriers. As for the FokI polymorphism, Jurutka et al. [VDR FokI polymorphism are still required. In addition, more studies are needed to confirm our results.A considerable weight of evidence supporting a role for VDR in urolithiasis was derived from GHS rats. Experimental studies using GHS rats showed increased levels of VDR in intestine, bones and kidneys ,39. MoreGHS rats . They deGHS rats . PreviouGHS rats . In addiGHS rats ,44. Therg et al.  showed tg et al. . Furtherg et al.  showed tg et al. , who alsg et al. . Ruggierg et al.  found tha et al.  suggestea et al. . ResultsFokI and TaqI polymorphisms and urolithiasis risk. However, we found ApaI and TaqI polymorphisms were significantly associated with urolithiasis risk. More case–control studies were included in our meta-analysis than Lin’s study. Therefore, our study may be more powerful and the conclusion might be more reliable. This reason might explain the difference between these two meta-analyses. In addition, our study had some advantages. First, Lin and coworkers tested multiple genetic models, including allelic comparison, a dominant model, and a recessive model. However, they did not correct for multiple comparisons or give biologic rationale for the choice of these genetic models. When the underlying genetic model is unknown, it is better to use pairwise comparisons of the three genotypes to find the best genetic model [VDR-hypercalciuria interactions on urolithiasis risk. Third, we explored the haplotype effect of ApaI and TaqI polymorphisms on the susceptibility to urolithiasis. Finally, the methodological issues for meta-analysis, such as, quality assessment was well investigated.Our meta-analysis included a total of 23 case–control studies with 3046 cases and 3206 controls, while a previous meta-analysis by Lin et al.  only incic model . We usedBsmI and FokI polymorphisms. Subgroup analysis was used to explore the sources of heterogeneity. After subgroup analysis by the calciuria level, the heterogeneity was effectively disappeared in hypercalciuric patients. Thus, it could be presumed that the relatively large heterogeneity mainly resulted from the calciuria level. In addition, funnel plot and Egger’s tests were used to find potential publication bias. No significant publication bias was detected.We should point out the importance of heterogeneity and publication bias, which might influence the results of meta-analysis. In our study, significant heterogeneity was found in the ApaI, BsmI, and TaqI polymorphisms and urolithiasis risk.Some limitations of our study should be addressed. First, the number of studies that were included in this meta-analysis was moderate. It was possible that some relevant published studies or unpublished studies with negative results were missed. Second, all of the studies were performed in Asians and Caucasians; thus, our results may be applicable only to these ethnic groups. Third, this study did not address gene-gene and gene-environment interactions, because insufficient information could be extracted from the original publications. Fourth, the data of the stone types and biochemical profiles was not showed in our meta-analysis because of insufficient information from the primary publications. Finally, we could not investigate the association between the haplotypes of the VDR polymorphisms and urolithiasis susceptibility. Our results indicated that the ApaI and TaqI polymorphisms were associated with the risk of urolithiasis. Haplotype analysis suggested that ApaI-TaqI haplotypes conferred the susceptibility to urolithiasis. Future large-scale studies with more ethnic groups are needed to validate our findings. Gene-gene and gene-environment interactions should also be considered in future studies.To our knowledge, this was the most comprehensive meta-analysis to assess the relationship between the The authors have declared that no competing interests exist.PZ and WN collected the literature data, developed the statistical model, carried out the software implementation and drafted the manuscript. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2350/14/104/prepub


C4, which varies among individuals, may determine the intrinsic strength of the classical complement pathway. Presuming a major role of complement as an effecter in peptide-mediated inflammation and phagocytosis, we hypothesized that C4 genetic diversity may partially explain the development of Graves' disease (GD) and the variation in its outcomes.Gene copy number of complement component C4 isotypes (C4A and C4B) genes were performed by quantitative real-time polymerase chain reaction analysis. Statistical comparison and identification of CNV of total C4, C4 isotypes (C4A and C4B) and C4 polymorphisms were estimated according to the occurrence of GD and its associated clinical features.A case-control study including 624 patients with GD and 160 healthy individuals were enrolled. CNV of C4, C4A and C4B genes, especially those with A2B2 polymorphism may associate with the development of GD . Although the distribution of copy number for total C4, C4 isotypes as well as C4 polymorphisms did not associate with the occurrence of goiter, nodular hyperplasia, GO and myxedema, <2 copies of C4A may associate with high risk toward vitiligo in patients with GD .Individuals with 4, 2, and 2 copies of These results may be further estimated for its clinical application on GD and the vitiligo in patients with GD. Graves' disease (GD) is an organ-specific autoimmune thyroid disease . It has A new paradigm in human genetics is high frequencies of interindividual variation in the copy number (CN) of specific genomic DNA segments. Copy number variation (CNV) loci often contain genes engaged in host-environment interactions, including those involved in immune functions, which results in susceptibility or resistance to autoimmune diseases -7, howevComplement component C4 (C4), located on chromosome 6q21.3, is encoded by 2 separate loci in the major histocompatibility complex class III region and derives 2 functionally distinct C4A and C4B isoforms [C4 genetic diversity may partially explain the development of GD as well as the variation in its outcomes. Here we investigated the polymorphic variants of C4 that correlate with predisposition to this disease.isoforms . The comisoforms . Presumi-34).A total of 624 patients (227 with GO and 397 without GO) with a confirmed diagnosis of GD and an appropriate control group with 160 healthy volunteers from China Medical University Hospital in Taiwan were enrolled and followed actively. All individuals provided informed consent as approved by the ethics committee of China Medical University Hospital. For the patients, diagnosis of GD and GO was followed the criteria set previously . Full meC4 gene dosage was assessed by quantitative real-time TaqMan® PCR analysis (Applied Biosystems) as described in the previously published protocols with some modification [C4A, and C4B , VIC-conjugated TaqMan® RNase P control reagents , 250 nM of the respective FAM-conjugated TaqMan® probes (C4A or C4B), the particular primers (300 nM C4A or C4B) in distilled water was contained in each of the distinct PCR batches. Appropriately prediluted genomic DNA  was added before start. CN of each target gene in each sample was determined from three separated experiments. Thermal cycler conditions were adjusted as follows: initial denaturation step for 10 minutes at 95°C; 40 cycles including denaturation for 15 seconds at 95 °C; and annealing/extension for 1 minute for 60°C. The data were analyzed using SDS 2.3 software (Applied Biosystems).Genomic DNA was extracted from peripheral blood following the manufactory's suggestions (Qiagen). fication . Real-tiT value of RNase P, C4A or C4B was converted into a raw gene dosage by the formula nRAWC4X = 2 RNase P)-(CTC4X)+1 (99% confidence for the simulation result). Odds ratios (ORs) and 95% CIs were estimated from logistic regression models adjusting for confounding variables as shown in Table Statistical analysis was performed using the statistical package PASW for Windows . The demographics of patients and healthy individuals were analyzed by the chi-square analysis. For those with 2 × 2 contingency tables, differences in the incidence of individuals with C4, C4 isotypes as well as C4 polymorphisms according to the presence of GD is shown in Table C4 alleles. After adjusting for age, individuals with 4 copies of C4 gene were more susceptible to GD  as compared to those without, whereas those with <4 copies of C4 gene tended to prevent from GD  as compared to those without. The distribution of C4A and C4B among individuals with or without GD was further investigated. For C4A gene, individuals with 2 copies of C4A increased the risk toward GD  whereas those with <2 copies of C4A reduced the risk toward GD . For C4B gene, individuals with 2 copies of C4B increased the risk toward GD  whereas those without 2 copies of C4B reduced the risk toward GD . Polymorphism analysis indicated tat individuals with the most common polymorphism (37.3%), A2B2, with 2.671-fold risk toward GD  as compared to those without. These results indicate that individuals with 4, 2 and 2 copies of C4, C4A and C4B genes, especially those with A2B2 polymorphism may have higher risk, whereas those with<4, <2 and ≠2 copies of C4, C4A and C4B genes may have lower risk toward GD, respectively.The distribution of copy number for total C4 genes and clinical features of GD. CNV of C4 genes showed association with susceptibility toward GO, vitiligo and myxedema, but not goiter or nodular hyperplasia as estimated by Fisher's exert test (data not shown). After adjusting for age, nodular hyperplasia, GO, and vitiligo, the distribution of copy number for total C4, C4 isotypes as well as C4 polymorphisms did not associate with the occurrence of myxedema . The distribution of C4A and C4B among GD patients with and without GO were further investigated. After adjusting for age, gender, radioiodine treatment, vitiligo and myxedema, neither isotypes nor polymorphisms of C4 was significantly associated with GO, although GD patients with <2 copies (0 or 1) of the C4A gene were less susceptible to GO  as compared to those with 2 copies of C4A, and those with A3B1 polymorphism were less susceptible to GO  as compared to those with A2B2 polymorphism. These results indicate that neither isotypes nor polymorphisms of C4 was significantly associated with GO, however, as compared to GD patients with 2 copies of C4A or those with A2B2 polymorphism, those with <2 copies of C4A or those with A3B1 might be protected against the development of GO, respectively.The distribution of copy number for total C4, C4 isotypes as well as C4 polymorphisms according to the presence of vitiligo is shown in Table C4A had a 5.153-fold increased risk of vitiligo . It remained significant even when compared with GD patients with 2 copies of C4A  and the SLE-related renal damage [C4A may enhance the development of vitiligo in GD patients, implying exist of an alternative pathway for the deficiency of complement.However, it may play a different regulatory role in systemic autoimmune diseases. Low level of C4 complements in sera has been found in several autoimmune diseases -21. In al damage ,19. A hyC4 CNV to GD as well as other GD clinical features, only the lower copies of C4A, but not C4B, were associated with higher risk of vitiligo. Because it appears that C4A binds to amino group-containing antigens such as immune complex, whereas C4B binds to hydroxyl group-containing antigens such as bacteria, this result may provide another view to support the hypotheses that the pathogenesis of vitiligo may be more relevant to the existence of the immune complex than the pathogen. In addition, recent studies have identified that the risk locus within the major histocompatibility complex region on chromosome 6q may be associated with vitiligo in both Chinese Han population and American population [C4 polymorphism and the vitiligo risky locus. Moreover, although confirmation of these results in larger samples is warranted, it would be interesting to further investigate the functional role of C4A in the development of vitiligo.What is interesting is that although we explored the relationship of pulation ,23. It mC4, C4A or C4B may associate with the development of GD and <2 copies of C4A may associate with development of vitiligo in patients with GD. These results may be further estimated for its application on predicting the occurrence of GD and the clinical outcome in patients with GD which might aid in the diagnosis of the disease and the development of therapeutic strategies.This study provides evidence that the CNV of (GD): Graves' disease; (GO): Graves' ophthalmopathy; (CNV): copy number variation; (CN): copy number; (SLE): systemic lupus erythematosus.The authors declare that they have no competing interests.YHL designed the study, managed the literature searches, undertook the statistical analysis, and wrote the draft of the manuscript. LW designed and performed the experiments. CTC and WCC recruited and maintained the clinical information of participants. LWLL and TYT undertook the statistical analysis. CHT and FJT directed the study and reviewed the results. All authors contributed to and have approved the final manuscript.


CCND1) plays a vital role in cancer cell cycle progression. Numerous epidemiological studies have evaluated the association between the CCND1 G870A polymorphism and the risk of colorectal cancer. However, these studies have yielded conflicting results. To derive a more precise estimation of this association, we conducted a meta-analysis and systematic review.Cyclin D1  with 95% confidence intervals (CIs) were derived from a fixed effect or random effect model. We applied a grading system (Venice criteria) that assessed the epidemiological strength of the association. A total of 22 publications that included 6157 cases and 8198 controls were identified. We found that the CCND1 G870A polymorphism was significantly associated with overall colorectal cancer risk . After further stratified analyses, the increased risk was observed only in the subgroups of hospital-based studies, PCR-RFLP genotyping methods, sporadic colorectal cancer, and Caucasian ethnicity.A comprehensive search was conducted to identify eligible studies of the CCND1 870A allele might be a low-penetrant risk factor for colorectal cancer.The available evidence demonstrates that the  Colorectal cancer (CRC) is the second most common type of cancer in women and the third most common type in men in the United States and Europe CCND1 G870A polymorphism, rs603965) and excessive cyclin D1 activity are common in numerous human tumors, including breast cancer, lung cancer, head and neck cancers, gastric cancer, gynecological cancers, blood-related cancers, and CRC CCND1 G870A polymorphism to increased CRC risk, the results remain controversial. To further investigate the combined effect of the CCND1 G870A polymorphism on CRC susceptibility, we performed a meta-analysis and systematic review.The adenine-to-guanine (A/G) substitution at nucleotide 870  the CCND1 G870A polymorphism was determined; (2) the outcome had to be colorectal cancer in humans. The major exclusion criteria were (1) reviews, tutorials, letters, and editorials; (2) duplicate data; (3) not a case-control design; (4) insufficient data were reported as cyclin D1 expression levels were provided without genotype data; (5) overlapping data and data superseded by the latest reports.All published literature investigating an association between the Data were extracted independently and crosschecked against the research consensus. The following variables were recorded: the first author’s last name; publication year; region/country where the study was performed; participant gender; ethnicity  of the study population; epidemiological type of colorectal cancer , sporadic colorectal cancer (sCRC), and sporadic colonic cancer (sCC)); histopathological subgroup information if known (included Dukes’ stage (A/B and C/D) and degree of differentiation ); control source (family-based study (FB), population-based study (PB), and hospital-based study (HB)); genotyping method (polymerase chain reaction (PCR) single-stranded conformation polymorphism (PCR-SSCP), PCR restriction fragment length polymorphism (PCR-RFLP), high-performance liquid chromatography (HPLC), TaqMan PCR, and DNA sequencing); sample size ; and the P value of the Hardy-Weinberg equilibrium in the control group. Only the latest studies were included when the data sets overlapped or were duplicated. The primary authors were contacted to provide additional information when necessary. Study identification and data extraction were conducted independently by three investigators and checked for accuracy by one author.Dichotomous variables were pooled using an odds ratio (OR). The summary OR was replaced by the risk difference (RD) if one of the studies reported no events in either the case group or the control group.The wild type G/G genotype was considered as a reference. Pooled effects were calculated for a homozygote comparison model (A/A vs. G/G), a heterozygote comparison model (G/A vs. G/G), a dominant model (G/A+A/A vs. G/G), and a recessive model (A/A vs. G/G+G/A).2 statistic, heterogeneity was defined as low or moderate if less than 50% and high if greater than 50% The statistical heterogeneity between included studies was determined using the chi-square-based Q-test Sensitivity analyses were conducted either by replacing a value of effect with another or removing individual studies from the data set. Sensitivity analyses were also performed by excluding studies in which the genotype frequencies in the controls significantly deviated from the HWE. We conducted subgroup analyses of the study design, cancer type, cancer location, ethnicity, Dukes’ stage, degree of differentiation, gender and genotyping method to investigate potential sources of heterogeneity.Publication bias among the included studies was assessed graphically using a Begg’s funnel plot The study confidence interval (CI) was established at 95%. Two-tailed P values of less than 0.05 were considered statistically significant. All statistical analyses were performed using the STATA version 11.0 software .2 statistic The Venice criteria CCND1 G870A polymorphism and colorectal cancer susceptibility were identified based on MOOSE  guidelines Through literature search and selection, a total of 22 publications Five articles 2 = 11.20) and the dominant genetic model , but heterogeneity was significant for the homozygote genetic model  and the recessive genetic model .The genotype data in the 22 studies were homogenous for the heterozygote genetic model , subsets of sCRC cases , subsets of Caucasian ethnicity , subsets of Duke’s stage C/D , subsets of the well/moderate degree of differentiation , male subjects , and subsets of the PCR-RFLP genotyping method . Specifically, the subgroup of Caucasian ethnicity was associated with 1.3- to 1.5-fold increased risk of sCRC without heterogeneity  (Significant association of the = 0.002) Table 2.Sensitivity analyses was performed by omitting one study at a time. This procedure did not influence the pooled value, which supports the robustness of this current meta-analysis.The Begg’s funnel plot and the Egger’s test  showed no evidence of publication bias Figure 4CCND1 G870A polymorphism and colorectal cancer susceptibility. The total sample size (6157 cases and 8198 controls) in our meta-analysis exceeded 1000. Therefore, we assigned the amount of evidence category an A grade. Next, we assessed the extent of replication. Our meta-analysis showed a significantly increased risk of colorectal cancer in the homozygote genetic model, the heterozygote genetic model and the dominant genetic model but not in the recessive model in any category. We observed minimal heterogeneity in the heterozygote genetic model and the dominant genetic model and moderate heterogeneity in the heterozygote genetic model. Therefore, we assigned a B grade for the extent of replication. Finally, there was no evidence of publication bias in our pooled data, and most of the included studies were well matched for race, ethnicity, gender and age. The summary ORs of each genetic model were greater than 1.15; therefore, bias could not have easily rendered the observed association. Nevertheless, most studies did not publish sufficient information about whether the G870A polymorphism was relevant to other polymorphisms or other candidate genes. Therefore, the Venice criterion of protection from bias was given a B grade. The overall grade of the Venice criteria for our data was “ABB”, which is consistent with moderate evidence demonstrating the linkage between the G870A polymorphism and colorectal cancer risk.We applied the Venice criteria CCND1 or PRAD1, located on 11q13) consists of five exons and four introns and encodes cyclin D, a key regulatory protein promoting the transition through the restriction point in the G1 phase CCND1 have been identified and cataloged in public SNP databases . Of the polymorphisms identified, the common adenine-to-guanine (A/G) substitution at nucleotide 870 in the conserved splice donor region of exon 4 has received the most investigation CCND1 G870A polymorphism at the boundary of exon 4 and intron 4 affects alternative splicing and results in an variant transcript for cyclin D1, termed cyclin D1b, which lacks exon 5 Cell cycle regulation plays an important role in the evolution of cancer by influencing cell proliferation, differentiation and apoptosis CCND1 G870A polymorphism with CRC risk. A total of 6157 cases and 8198 controls were included. Based on the Venice criteria, the results indicated that the G/A or A/A genotype of CCND1 SNP rs603965 was significantly associated with an increased risk of CRC. Additionally, we found no significant risk of CRC associated with the CCND1 G870A polymorphism for the recessive model in any category, indirectly suggesting the linkage of the A-allele and increased CRC risk.The current meta-analysis and systematic review summarizes the results from 22 case-control studies on the association of the CCND1 G870A polymorphism and CRC risk remained significant in Caucasians and sCRC but not in Asians or HNPCC, which supports the hypothesis that genetic backgrounds and the environment in which patients live in might play important roles in the development of CRC CCND1 genotype and CRC risk was observed in the comparison model of either the colon subgroup or the rectum subgroup was in contrast with the results from another meta-analysis investigating digestive tract cancers and the risk associated with the CCND1 G870A polymorphism In the stratified analyses, the results showed that the association between the Meta-analysis is an important tool for revealing trends that might not be apparent in a single study. The pooling of independent but similar studies increases precision and therefore increases the confidence level of the findings. The current meta-analysis has some advantages. First, the number of total cases and controls was substantial, which significantly increased the statistical power of the analysis. Second, no publication biases were detected, which indicates that the entire pooled result may be unbiased.CCND1 G870A polymorphism is independently predictive of cancer risk remains controversial CCND1 G870A and caspase−8 6 n del/ins on CRC CCND1 for colorectal cancer, although one group recently published a GWAS in which CCND1 was strongly suggestive in melanoma carcinogenesis CCND1 G870A polymorphism with CRC risk.Despite these advantages, some limitations in the current meta-analysis should be acknowledged. First, the controls were not uniformly defined. Although most of the patients in the control groups were selected from healthy populations, some might have had a benign disease. Therefore, there was a lack of proper matching, and the results are based on unadjusted estimates. The current meta-analysis is unable to solve problems with confounding factors that could be inherent in the included studies. Inadequate control of the confounders might bias the results either toward exaggeration or underestimation of risk estimates. Second, stratifying analyses were based on a relatively small number of studies from which detailed individual data were available; therefore, some of the subgroup analyses were difficult to perform. Third, although there is no indication of major publication bias in the formal evaluation used, potential publication bias is impossible to completely exclude because small studies with null results tend to not be published. Finally and mostly importantly, whether the CCND1 G870A polymorphism is associated with CRC susceptibility, especially among patients of Caucasian ethnicity. The current results may prompt further investigation of diagnostic approaches and prevention strategies to combat CRC.In conclusion, the current meta-analysis and systematic review demonstrated that the


Transforming growth factor-beta (TGF-β1) gene has been regarded as an important mechanism in angiogenesis, endothelial cell proliferation, adhesion,and the deposition of extracellular matrix. The TGF-β1 gene may be involved in the development of diabetic retinopathy (DR) through disrupting angiogenesis. However, studies investigating the relationship between −509C/T and +869T/C(L10P) polymorphisms and DR yielded contradictory and inconclusive outcomes. In order to realize these ambiguous findings, a meta-analysis was performed to assess the association between the TGF-β1 gene polymorphisms and susceptibility to DR.We conducted a search of all English reports on studies for the association between the TGF-β1 gene polymorphisms and susceptibility to DR using Medline, the Cochrane Library, EMbase, Web of Science, Google (scholar), and all Chinese reports were identified manually and on-line using CBMDisc, Chongqing VIP database, and CNKI database. The strict selection criteria and exclusion criteria were determined, and odds ratios (ORs) with 95% confidence intervals (CIs) were used to assess the strength of associations. The fixed or random effect model was selected based on the heterogeneity test among studies. Publication bias was estimated using Begg's funnel plots and Egger's regression test.A total of three studies were included in the meta-analysis and all included studies analyzed patients with type 2 diabetes. For +869T/C(L10P) polymorphism, significant association was observed in an allele model  and the recessive model . As regards −509C/T polymorphism, no obvious associations were found for all genetic models.This meta-analysis suggested that the +869T/C(L10P) polymorphism in TGFβ1 gene would be a potential protect factor for DR. However, the −509C/T polymorphism is not associated with DR. It has been widely accepted that diabetic retinopathy (DR) is one of the foremost causes of blindness in the working age population TGF-β1 has an important role in angiogenesis, endothelial cell proliferation, adhesion and the deposition of extracellular matrix Thus far, previous studies concerning association between −509C/T polymorphism and risk of DR are limited and rather conflicting within type 2 diabetes Two reviewers (Lei Liu and Jinghua Jiao) independently scrutinized studies on the associations between TGFβ1 gene polymorphisms and DR. We searched Medline , Embase , the Cochrane Library , Chinese Biological Medicine , China National Knowledge Infrastructure , Wang Fang Data  and Chongqing VIP database  databases using the terms “transforming growth factor beta 1 or transforming growth factorβ1 or TGF beta 1 or TGFβ1”, “diabetes or diabetic complications” and “polymorphism, variant or mutation”. We used the PubMed option “Related Articles” for each study to retrieve additional potentially relevant articles. Reference lists were checked and researchers contacted for additional literatures. Authors of publications were contacted when results were unclear or when relevant data were not reported. The search was done without restriction on language, but we only included published articles written in English or Chinese.Studies were selected if they met the following criteria: (1) association study in sporadic DR subjects; (2) there were available data for TGFβ1 gene mutations with risk of DR, using a case-control or cohort design; (3) the genotype distribution in the controls of all studies should be in agreement with Hardy-Weinberg equilibrium (HWE); and (4) in the case of multiple publications from the same study group, the most complete and recent results were used.The exclusion criteria were defined as: 1) useless data reported, genotype number or frequency not included; 2) abstracts, reviews and animal studies; and 3) genotype distribution in the control population not consistent with HWE.P value for HWE.After excluding the overlap studies and including the additional ones, two investigators independently extracted data from each study with a standard fashion and entered into a common database. When discrepancies were appeared, all investigators were recruited to assess the data. The following information was collected: First author, year of publication, country, ethnicity, characteristics, study design, sample sizes of patients and controls, genotype numbers, minor allele frequency (MAF) and numbers (MAN), The review and analysis were guided to conduct by the PRISMA statement for preferred reporting of meta-analysis 2I statistic interpreted as the proportion of total variation contributed by between-study variation. If there was a statistical difference in terms of heterogeneity, the random effects model would be used to estimate the pooled ORs P values were two sided and a P value of smaller than 0.05 for any test was considered to be statistically significant.Odds ratio (ORs) with 95% confidence intervals (CIs) for genotypes and alleles were used to assess the strength of association between TGFβ1 gene polymorphisms and DR. The ORs were performed for the allele contrasts, additive genetic model, as well as recessive genetic model and dominant genetic model, respectively. Heterogeneity was examined with Finally, a total of three studies fulfilling the inclusion criteria were identified As shown in The association between −509C/T polymorphism and DR were also shown in There was no significant publication bias according to Begg's and Egger's tests .In this study, we deleted every single study each time, in order to examining the influence of the individual data set to the pooled ORs. According to sensitivity analytical results, we found that there was no substantial modification of our estimates after exclusion of individual studies, indicating that the results were stable (data not shown).There are three TGF-β isoforms, TGFβ1-3, which play important roles in regulating inflammation, cell growth and differentiation Previous research suggested that +869T/C(L10P) gene polymorphism is associated with diabetic nephropathy in Chinese. The role of TGFβ1 in pathogenesis of diabetic nephropathy is pathologic tissue fibrosis leading to organ failure Except for evaluating the expression of TGFβ1 in vivo or in vitro To the best of our knowledge, this is the first meta-analysis to explore TGFβ1 polymorphism in development and progression of DR. To achieve a more reliable and comprehensive conclusion on both variants, we used meta-analysis to assess the association between the TGFβ1 gene and DR risk on the basis of data from three studies. We summarized a significant association for +869T/C(L10P) polymorphism, especially in an allele model  and the recessive model . Because the included studies were case-control studies, we did not perform a subgroup analysis by study design. However, the results indicated no relationship of −509C/T polymorphism with DR risk . The results from our study seem to indicate that the −509C/T polymorphism may be not associated with DR risk. Because heterogeneity was found among the studies, we employed random-effect model. Then a sensitivity analysis was carried out by removing one study for each time and re-running the model to determine the effect on the overall estimate. The estimates changed quite little, indicating that the results were stable. Owing to the limited number of included studies, we did not perform a publication bias test.In addition, some other TGFβ1 genes including −988C/A, −800G/A, and +915G/C(R25P) were studied in the relationship with DR. The −988A allele was not represented in the study and no signicant differences between groups were found for the −800G/A. However, +915G/C(R25P) polymorphisms in the TGF-β1 gene could be regarded as a strong genetic risk factor for DR In this meta-analysis, all included studies analyzed patients with type 2 diabetes. The etiology and mechanisms of type 1 and 2 diabetes are different, so there may be diversity in the association between TGFβ1 gene polymorphism and DR with type 1 or 2 diabetes. However, the relationship between TGFβ1 gene polymorphism and DR within type 1 diabetes is not demonstrated. More efforts are encouraged to explore this association.This meta-analysis has pooled all the available results from the case-control studies, which has significantly increased the statistical power. However, the results of the present meta-analysis should also be interpreted within the context of its limitations. First, DR is a multi-factorial disease from complex interactions between environmental exposures and genes factors. In this meta-analysis, we had insufficient data to perform an evaluation of such interactions for the independent role of TGFβ1 polymorphisms in DR development. Second, the major limitation of our study is the relatively limited available studies included in the meta-analysis. Thus, investigations involving large size of different races are necessary for a more reliable evaluation on their associations. Third, our meta-analysis is based on unadjusted estimates because of a lack of original data. Forth, although every effort was made to ascertain all appropriate publications, it is likely that some were missed or displayed erroneously. In addition, we did not consider studies published in languages other than English/Chinese or data presented in abstracted form.In spite of these limitations, for DR candidate genes, we believe that the positive locus identified in our systematic meta-analyses that warrant follow-up with high priority with a prospective trial.In conclusion, this meta-analysis suggested that the +869T/C(L10P) polymorphisms in the TGF-β1 gene could be regarded as a strong genetic risk factor for DR. However, the −509T/C polymorphism is not associated with DR risk. At the same time, this result should be interpreted cautiously. To better understand the potential mechanism for DR in humans, in the future, large well-designed epidemiological studies in the susceptibility of DR evidence are needed to confirm this association. It also will be necessary to combine genetic factors and other environmental risk factors.Checklist S1PRISMA 2009 Checklist.(DOC)Click here for additional data file.


The study polymorphism was selected based on recent reports and a literature meta-analysis suggesting association with increased risk of various types of cancer. We screened 324 AChR+ MG patients  and 735 healthy matched controls . 94 of the total MG patients had a thymoma. While there was no association with the whole cohort of MG patients, we found a statistically significant association of the DNMT3B-579T allele  and the TT homozygous genotype  with the risk of thymoma. No association was observed in MG patients without thymoma, even after stratification into clinical subtypes. Present results suggest that the DNMT3B-579T allele might contribute to the risk of developing thymoma in MG patients, particularly in homozygous TT subjects.Increasing evidence suggests a contribution of epigenetic processes in promoting cancer and autoimmunity. Myasthenia gravis (MG) is an autoimmune disease mediated, in approximately 80% of the patients, by antibodies against the nicotinic acetylcholine receptor (AChR+). Moreover, epithelial tumours (thymomas) are present in about 10-20% of the patients, and there is indication that changes in DNA methylation might contribute to the risk and progression of thymomas. However, the role of epigenetics in MG is still not completely clarified. In the present study we investigated if a common polymorphism ( Increasing evidence suggests a contribution of epigenetic modifications in complex diseases such as cancer and autoimmune disorders -3. The tS-adenosylmethionine (SAM) as the methyl donor compound [DNA methylation represents one of the most studied epigenetic marks for gene regulation; it consists of the addition of a methyl group to the 5’ position of the cytosine pyrimidine ring (5-methylcytosine) mediated by DNA methyltransferase enzymes (DNMTs) using compound . Methylacompound . Most ofcompound . There icompound ,7,8.MGMT, hMLH1, p16/INK4, and RASSF1A, whose increased methylation significantly correlated with tumour severity [Myasthenia Gravis (MG) is a prototypic antibody-mediated autoimmune disease. In approximately 80% of the patients the disease is mediated by auto-antibodies against the nicotinic acetylcholine receptor (AChR) . Despiteseverity ,13.-579G>T: rs1569686) in the promoter of the DNMT3B gene coding for the DNA methyltransferase 3B, one of the key de novo enzymes mediating DNA methylation, increases the risk to develop MG or MG-associated thymomas. The selected polymorphism has been suggested to impair DNA methylation of tumor suppressor genes, therefore impairing their expression levels, and for this reason it was investigated as a susceptibility factor for several cancer types [DNMT3B promoter polymorphisms have been associated with global DNA methylation and are increasingly investigated in other complex diseases than cancer [The goal of the present study was to investigate if a common polymorphism  of the Pisa University Hospital, at the Institute of General Pathology of the Catholic University of Rome, and at the Department of Translational Research and New Technologies in Medicine and Surgery of the University of Pisa. Disease diagnosis was based on characteristic signs and symptoms of MG coupled with an anti-AChR antibody positive test; patients with an anti-AChR antibody negative test were excluded from the present study. The main clinical characteristics of MG patients are listed in 5’-GAG GTC TCA TTA TGC CTA GG-3’ and reverse: 5’-GGG AGC TCA CCT TCT AGA AA-3’), 0.15mM of each dNTPs, 1.5mM MgCl2, and 40 ng of genomic DNA in a total volume of 25µl. PCR conditions were 35 cycles of 30s at 95°C, 45s at 56°C, and 45s at 72°C, preceded by an initial denaturation step of 5 min at 95°C and followed by a final elongation step of 10 min at 72°C. Overnight digestion at 37°C with Pvu II  resulted in an undigested 225-bp product for the wild type allele (G), and in 132- and 93-bp products for the mutant allele (T). Digestion products were visualized after electrophoresis on a 2% agarose gel containing ethidium bromide. Samples with previously confirmed wild-type, heterozygous and mutant genotypes [Genomic DNA was extracted from peripheral blood lymphocytes by means of the QIAamp® Blood Mini Kit  according to the manufacturer’s protocol. Genotyping analysis was performed by means of PCR-RFLP techniques following the protocol of Lee et al.  with minenotypes  have bee2) analysis. Differences in allele and genotype distributions between groups have been evaluated by means of Fisher’s exact test. Odds ratios (ORs) have been calculated by means of unconditional logistic regression and given with 95% confidence intervals (CIs). All individual values were analyzed with the SPSS 16.0 statistical package for Windows. The statistical power of the study was evaluated by means of the statistical package QUANTO 1.2.4.exe. Given a case-control cohort of at least 300 cases and 700 controls, and a minor allele frequency (MAF) of 31% in Controls, the study had an a priory power of 82% to detect differences in allele frequencies.Differences in mean age and gender between groups were evaluated by means of the Student’s T-test, and Fisher’s exact test, respectively. To verify that genotype frequencies were in Hardy-Weinberg equilibrium in controls we used Chi-square (χDNMT3B-579G>T polymorphism in patients and controls are shown in P = 0.18). No difference in allele and genotype frequencies was observed between patients and controls when MG patients were considered as a whole . Moreover, the homozygous TT genotype was associated with an increased thymoma risk compared to the homozygous GG one . Both associations remained significant even after Bonferroni’s correction for multiple comparisons (Allele and genotype frequencies generated by the  a whole  and 3. Aparisons  and 3. GDNMT3B-579G>T polymorphism in MG patients stratified according to disease age at onset (≤45 years vs. > 45 years), to disease severity (less severe forms: Grade I + IIA vs. more severe forms: Grade IIB + III + IV), or to the presence/absence of associated autoimmune diseases (associated AID vs. no associated AID). No difference in allele and genotype frequencies was observed in young onset vs. late onset MG patients, in less severe vs. more severe MG, or in patients with associated AID with respect to those with no associated AID (data not shown). We also compared allele and genotype frequencies generated by the DNMT3B-579G>T polymorphism in a large population of MG patients of Italian descent. Overall, no association of the studied polymorphism with MG was observed, even after stratification of the patients according to different clinical parameters like age at onset, disease severity according to Osserman classification, or presence/absence of other autoimmune diseases. However, when we stratified MG patients according to thymus pathology, we found a statistically significant association of the T allele with the presence of thymoma, particularly among carriers of the homozygous TT genotype. In order to shed some light on the role of epigenetics in both MG and MG-associated diseases, we analyzed the DNMT3B-579T allele [DNMT3B-579G allele in Asians is much lower than that of the DNMT3B-579T one [Present data are in agreement with a literature meta-analysis suggesting an increased risk of cancer associated with the presence of the T allele . That meT allele . Interes579T one . However579T one -21, but hMLH1, RASSF1A, MGMT, p16/INK4, DAPK, FHIT, RAR2, CDH1, and APC, are frequently hypermethylated in thymic epithelial tumours, all found to be methylated in almost 20 to 40% of the cases [Hirose and coworkers  investighe cases ,30. Sevehe cases . DNMT3B-579G>T polymorphism is not yet completely elucidated. Some authors suggest that it can directly impair promoter activity and gene expression levels, and others observed a linkage disequilibrium between rs1569686 and other DNMT3B promoter polymorphisms, namely −149C>T (rs2424913) and -283T>C (rs6058870), which have been functionally associated with promoter activity and gene expression levels [DNMT3B-579G>T polymorphism with various types of cancer, suggest that this polymorphism might either have a functional role on gene expression levels, or be a tag SNP of functional haplotypes [DNMT3B promoter polymorphisms, DNMT3B protein levels, and DNA methylation levels, suggesting that the increased risk of cancer observed in carriers of those polymorphisms might be due to impairments of DNA methylation in cancer cells [The functional role of the n levels ,25,28,31plotypes ,28,31. Ier cells ,32. Except the presence of thymoma, no other investigated MG clinical characteristic was associated with this polymorphism in our population. Interestingly, others evaluated the contribution of this polymorphism to the onset and progression of autoimmune thyroid disease, and found no association . The obsDNMT3B promoter polymorphisms in MG patients. Our study suggests that the presence of the DNMT3B-579T allele might represent a risk factor for the development of MG-associated thymomas, particularly in carriers of the homozygous TT genotype, while it probably does not have effect on other MG subtypes. Therefore, the analysis of this polymorphism could help to identify those AChR+ MG individuals at increased risk to develop a thymoma.The thymus plays distinct roles in the pathogenesis of the different AChR+ MG subtypes. Inflammatory, neoplastic and age-related alterations of the thymus are of pivotal relevance for the initiation of anti AChR autoimmunity in early onset MG, thymoma-associated MG and, likely, late onset MG, respectively . At best