Hydrofolate Reductase Research Articles

Detection of C677T, rs75582903 and SNV64878785 in Methylenetetra hydrofolate Reductase Gene (MTHFR) in Meningioma among Sudanese patients

Introduction: Meningiomas are commonly benign tumors come from the arachnoid cap cells, and the incidence is account 6 cases per 1000000 of population per year or even much higher in African countries, in Sudan during the period May 2005-May 2012 a total of 405 patients were operated upon and diagnosed as having cranial meningioma, regarding the etiologies of meningiomas are not yet known; it may be related to radiation exposure, genetic disorders (Neurofibromatosis type2), hormonal imbalance (progesterone) and single nucleotide polymorphisms (SNPs).
 Aims of the study: This study aimed to amplify MTHFR gene in specific region using PCR, and to detect C677T, rs755829023 and SNV64878785 of MTHFR gene in meningioma among Sudanese patients.
 Materials and Methods: This was a cross-sectional study conducted at the National Center for Neurological Sciences between May to August 2022, Khartoum Sudan, all meningioma patients attending the national center of neurological sciences during the period of the study was included. Demographic variables were included (age and gender), and the clinical data included (meningioma histology subtype and WHO grading), tissue and blood samples were collected, DNA extraction from the tissue and blood samples was done, polymerase chain reaction was made to amplify specific region of MTHFR gene. PCR products were sent for Sanger sequencing to Macrogen Europe Laboratory. Data was collected and analyzed by using Statistical Package for the Social Sciences (SPSS) version 19, and sequencing was analyzed using, BioEdit and Mutation taster.
 Results: In this study the most affected age group was 41-50 years, female was the most common, WHO grade I meningioma constituted 70% of the cases mostly the fibrous subtype. C>T (g.10369 C>T) was detected in 100% of the cases and deletion C (g.10380-10380delC) was detected only in the cases 16.6%. 
 Conclusion: In this study, female was the most common 11(55%) with male to female ratio 1:2, the most affected age group was ranging from 41-50 years constituted 35%, WHO grade I meningioma was detected in 70% of the patients. C>T (g.10369 C>T) was detected in 100% of the cases and deletion C (g.10380-10380delC) was detected only in the cases 16.6%. 

MTHFR C677T and TS 5’-UTR 3R/3R Gene Polymorphism in Methotrexate-Resistant Childhood Acute Lymphoblastic Leukemia

BACKGROUND: Childhood acute lymphoblastic leukemia (ALL) is the most common pediatric malignancy in Indonesia and often treated by methotrexate (MTX). Though it can be cured in 30-60% of patients, MTX resistance remains the major cause of treatment failure in childhood ALL. Previous sudies showed that its anti-leukemic property was moderated by MTX ability to inhibitmethylene tetra hydrofolate reductase (MTHFR) and thymidylate synthase (TS) in folate metabolism. This study investigates the correlation between MTHFR and TS polymorphism and MTX resistance in ALL children.METHODS: A total of 155 subjects obtained from all subjects prior to chemotherapy. DNA from blood samples were extracted and underwent polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) to evaluate MTHFR C677T and TS 5’-UTR 3R/3R polymorphism.RESULTS: There was significant correlation between MTHFR C677T and TS 5’-UTR 3R/3R gene polymorphism with MTX resistance. Subjectswith MTHFR C677T and TS 5’-UTR 3R/3R gene polymorphism were 4 times (p=0.007) and 6.4 times (p=0.001) more likely to be MTX resistant than those without gene polymorphisms, respectively.CONCLUSION: MTHFR C677T andTS 5’-UTR 3R/3R represent dominant gene polymorphism related to MTX resistance in childhood ALL.KEYWORDS: gene polymorphism, folate metabolism, acute lymphoblastic leukemia

Polymorphism gene methylene tetra hydrofolate reductase (MTHFR) C677T and recurrence within one month end point in ischaemic stroke patients with and without hypertension

Background The genetic mutations associated with stroke are multiple and work in combination rather than isolated genes. Although many studies have emphasized strategies for the primary prevention of stroke, the prevention of recurrent stroke has received less attention. Recurrent stroke occurs most frequently in the early period after ischaemic stroke, with estimated ranging from 1,2%–9%. Aim This study aimed to know about the association between polymorphism gene MTHFR C677T and recurrence within one month in ischaemic stroke patients with and without hypertension. Material and methods The study was conducted with the approval of the Ethic Committee of the institutions involved and written informed consent was obtained from all subjects or their relatives (for comatose patients). All sample matched with inclusion and exclusion criteria. The neurological deficits confirmed in all cases by computerized tomography (CT) scan. We chose the outcome of recurrence within one month. Results A total of 106 were recruited, comprising 53 cases ischaemic stroke with hypertension 53 cases without hypertension. We have done the PCR- RFLP to all the patients, we got 78 patients with 677CC of MTHFR genotype, 23 patients with 677CT genotype and 5 patients with 677TT genotype. The association between polymorphism gen MTHFR C677T and recurrence was not significant but we found that recurrence within one month in genotype TT was the highest (20%). Conclusion We assumed that polymorphism MTHFR C677T have an important role with recurrence in early period after ischaemic stroke.

The Role of Polymorphism Gen Methylene Tetra Hydrofolate Reductase (MTHFR) C677T in Ischaemic Stroke Patients with and Without Hypertension.

BACKGROUND:Stroke is a leading cause of disability and remains the second leading cause of death in the world. Some of the pathogenesis of stroke are interactions between genetic and acquired risk factors, the interaction is related with the atherosclerotic which is the main pathogenesis of ischaemic stroke. Previous studies demonstrated an association between methylene tetra hydro folate reductase (MTHFR) genotype and ischaemic stroke; the MTHFR C677T genotype is one of the independent risk factor.AIM:This study aims to know about the role of polymorphism gen MTHFR C677T in ischaemic stroke patients with and without hypertension.METHODS:This study is a cross-sectional study, the sample was taken consecutively, after approval by the Medical Faculty Science’s Ethics Committee at University Sumatera Utara. All sample matched with inclusion and exclusion criteria, demography data and blood sample were taken. Demography data were analysed using descriptive statistic.RESULTS:Of the 106 ischaemic stroke patients were divided into two groups, the first group is patients with hypertension (53 patients), and the second group is without hypertension (53 patients). We have done the PCR- RFLP to all the patients, we got 78 patients with 677CC of MTHFR genotype, 23 patients with 677CT genotype and 5 patients with 677TT genotype. We found polymorphism C677T is more frequent in ischaemic stroke patients with hypertension (16 patients; 69.5%), and all the patient with 677TT genotype are an ischaemic stroke with hypertension (5 patients; 100%).CONCLUSION:We concluded that polymorphism MTHFR C677T have an important role in hypertension and ischaemic stroke.

Association of plasma homocysteine, folic acid levels, and C677T polymorphism in methylene tetra hydrofolate reductase with risk of preeclampsia: a case-control study in Iranian women

Introduction: Preeclampsia is the most dangerous hypertension with unknown etiology. Preeclampsia is a kind of pregnancy-specific syndrome. Objective: The aim of this study was to evaluate the correlation between C677T polymorphism of MTHFR gene, folic acid, and homocysteine serum levels in Iranian pregnant women with preeclampsia. Materials and Methods: This study was performed in 129 pre-eclamptic pregnant women and 125 control individuals and MTHFR gene (C677T polymorphism) was determined by PCRRFLP method, and the plasma levels of the homocysteine and acid folic was measured by ELISA method. Result: The CC, CT, and TT genotypes were not significantly different in patients compared to control (p = 0.614). Low mean levels of homocysteine and folic acid in the preeclamptic cases were observed compared to control group. The levels of BMI, gestational age, and neonatal weight were statistically different in two groups and other variables revealed no significant difference between these groups. Conclusion: These findings showed that there was no correlation between the C677T polymorphism of MTHFR gene and preeclampsia but the TT genotype of C677T polymorphism seems to be a protective factor for preeclampsia. It is also concluded that in this study, homocysteine and folic acid serum levels and BMI are significantly affected in patients with preeclampsia compared to controls and can increase the risk of developing sever side effect to mothers and neonates.

MTHFR isoform carriers. 5-MTHF (5-methyl tetrahydrofolate) vs folic acid: a key to pregnancy outcome: a case series.

To evaluate the possibility of correcting metabolic defects in gametes and embryos due to methylene tetra hydrofolate reductase (MTHFR) isoforms C677T and A1298C, by supplementation with 5-methyl THF instead of synthetic folic acid. In these couples, high doses of folic acid lead to UMFA (un-metabolized folic acid) syndrome. Thirty couples with fertility problems lasting for at least 4years, such as recurrent fetal loss, premature ovarian insufficiency, or abnormal sperm parameters, with two thirds of them having failed assisted reproductive technology (ART) attempts were included in this program. For all couples, at least one of the partners was a carrier of one of the two main MTHFR isoforms. Most of the women had been previously treated unsuccessfully with high doses of folic acid (5mg/day), according to what is currently proposed in the literature. The couples carrying one of the isoforms were treated for 4months with 5-MTHF, at a dose of 600 micrograms per day, before attempting conception or starting another attempt at ART. The duration of treatment corresponding to an entire cycle of spermatogenesis is approximately 74days. In this first series of 33 couples, one couple was not followed-up, and two are still currently under treatment. No adverse effects were observed. Thirteen of the couples conceived spontaneously, the rest needing ART treatment in order to achieve pregnancy. Only three couples have, so far, not succeeded. The conventional use of large doses of folic acid (5mg/day) has become obsolete. Regular doses of folic acid (100-200μg) can be tolerated in the general population but should be abandoned in the presence of MTHFR mutations, as the biochemical/genetic background of the patient precludes a correct supply of 5-MTHF, the active compound. A physiological dose of 5-MTHF (800μg) bypasses the MTHFR block and is suggested to be an effective treatment for these couples. Moreover, it avoids potential adverse effects of the UMFA syndrome, which is suspected of causing immune dysfunction and other adverse pathological effects such as cancer (especially colorectal and prostate).

Meta-analysis study to evaluate the association of MTHFR C677T polymorphism with risk of ischemic stroke.

Ischemic stroke is a condition characterized by reduced blood supply to part of the brain, initiating the ischemic cascade, leading to dysfunction of the brain tissue in that area. It is one of the leading causes of death and disability and is estimated to cause around 5.7 million deaths worldwide. Methyl tetra hydro-folate reductase (MTHFR) is a rate limiting enzyme in the methyl cycle which catalyzes the only biochemical reaction which produces 5, Methyl tetra hydro folate, the co-substrate for the re-methylation of homocystiene to produce methionine. MTFHR C677T is a common mutation of MTHFR and those homozygous for the MTFHR C677T produce a thermo-labile form of the protein with drastically reduced catalytic activity resulting in elevated plasma homocystiene levels - a common risk factor for cardiovascular diseases. However, the role of MTHFR C677T in ischemic stroke remains unclear. To evaluate this association, we carried out a meta-analysis of existing published studies, which included 72 studies involving 12390 cases and 16274 controls. The forest plot was made to evaluate the overall risk of the mutation in the etiology of Ischemic Stroke. The overall Odds- ratio of the study was found to be 1.319 for random effects model, revealing a ∼32% increased risk of Ischemic stroke in the presence of MTHFR C667T mutation compared to controls. Publication bias in the study was analyzed using funnel plot which revealed that only 7 studies out of the 72 contributed to publication bias. These 7 studies were excluded and Meta-analysis was repeated for 65 studies and overall odds-ratio was 1.306, which showed that there was a 30% higher risk of Ischemic stroke in the presence of MTHFR C667T.

Stroke Due to Hypercoagulable State Can Mimic Multiple Sclerosis: A Case Report

Introduction: Stroke is the second major cause of mortality worldwide and in several cases, and it may lead to disability. Factor V Leiden is a common genetic thrombophilia, which causes activated protein C (APC) resistance. Hyperhomocysteinemia and factor V Leiden deficiency, two independent coagulopathy factors, can lead to venous and arterial infarctions in multiple small and large arteries and veins anywhere in the body. Case Report: Here, we report a unique case in which both hyperhomocysteinemia and factor V Leiden deficiency are documented together with MTHFR (C677T) (Methylene Tetra Hydro Folate Reductase) gene polymorphism and activated protein C resistance respectively. Conclusion: More interestingly, the mode of presentation in this case highly resembled that of progressive multiple sclerosis; all signs and symptoms slowly progressed without any systemic signs at first few years. Further studies needed to assess current outcomes.

Three Coagulation Related Mutations and Increased Risk of Myoma in Women of Fars Province

Background: Myoma is one of the most common benign tumor in the women’s genital tract and causing some implications such as miscarriages, anemia or even infertility. Since tumorigenesis is associated with a hypercoagulant state, we investigate the role of coagulation related variations in methylene-tetra hydrofolate reductase (MTHFR) and endothelial protein C receptor (EPCR) genes in patients with uterine myoma. Methods: In this case- control study, genotyping was performed for rs867186, rs1801131 and rs1801133 by amplification-refractory mutation system- polymerase chain reaction (ARMS-PCR) method in 73 patients with myoma and 73 healthy women. Data was analyzed by logistic regression and χ2 test through software SPSS 16. Results: A statistically significant difference was observed between patients and the control group regarding the frequency of allele C in the MTHFR (A1298C) gene (P = 0.01). The frequency of the genotypes carries at least one C allele (CC + AC) is higher in patients (13.7%) compared to controls (2.7%). This means the C allele increases the risk of the disease (P = 0.01). Although the frequency of the T allele in patients was higher than controls, no significant relationship was observed between it and the risk of the disease. Our studied population was completely homogeneous for allele A at EPCR locus. Conclusions: In conclusion, results revealed a significant association between A1298T variation and myoma incidence, but more research is needed to verify the role of this mutation as a genetic marker in uterine myoma.

Abstract A47: Meta-analysis of MTHFR polymorphisms in lung cancer: Population health and mutations in the world

Abstract Purpose: The purpose of this meta-analysis is to identify the associations of methylene-tetra- hydrofolate reductase (MTHFR) gene mutations and smoking as risk factors for lung cancer (LC). LC is the leading cause of cancer-related death worldwide. To-date, population health in relations to the epigenetics pathways is not clearly understood. Inconsistent findings have been cumulated to-date on the case control studies of MTHFR gene mutations in LC around the world. Procedure: On-line databases were searched from PubMed (PM) and PM Central databases for meta-analyses by using a combination of key words including LC, MTHFR, and case control studies. Additional studies were located by cross reference method from located studies. For this analysis, 22 studies published within the last 17 years were identified, containing both control and LC cases of human samples. These papers were reviewed and evaluated using the criteria for the quality of studies by two raters. Relative risks (RR) were calculated for increased or decreased risks for LC. New Data Findings: Preliminary analyses included 11799 cases and 12322 controls associating MTHFR677 locus and LC in 22 studies; 6916 cases and 8526 controls for MTHFR1298 locus in 14 studies. MTHFR677 TT variant as homozygous mutation type was associated with increased LC risk for all race groups combined (pooled RR = 1.151, 95% CI = 1.045 - 1.267, p = 0.004); and also significant for subgroups of Asians (14 studies, RR = 1.080, 1.007 - 1.158, p = 0.031) and Caucasians (8 studies, RR = 1.334, 1.017-1.749, p = 0.037). Asians combined had greater mutation rates on MTHFR677 TT and TC (TT+TC=19.6+47.2=66.8% for cases and 17.8+47.3=65.1% for controls) than Caucasians (11.4+43.4=54.8% for cases and 9.2+41.9%=51.1% for controls). For association of smoking and LC risk, pooled results from 12639 cases and 11828 controls in 17 studies associating MTHFR gene and LC indicated that smoking increased LC risk (pooled RR=1.116, 1.072 - 1.162, P < 0.001). Conclusion: Meta-analyses of epigenetic risk factors associating gene mutations and health behaviors can provide useful insights to improve the quality of future cancer prevention and intervention studies. Further studies are needed to document population-based gene mutations in relation to epigenetics pathways for cancer prevention. Citation Format: Hsiao-Ling Chen, Shyang-Yun Pamela K. Shiao, Linda H. Chiang, Joyce Kusuma. Meta-analysis of MTHFR polymorphisms in lung cancer: Population health and mutations in the world. [abstract]. In: Proceedings of the Thirteenth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2014 Sep 27-Oct 1; New Orleans, LA. Philadelphia (PA): AACR; Can Prev Res 2015;8(10 Suppl): Abstract nr A47.

Multi- analytical approach: better predictor of pharmacogenetic based clinical outcomes in breast cancer therapies

Background Chemotherapeutic drug clinical outcomes are genetically determined as there is a large heterogeneity in the response to, and toxicity of, chemotherapeutic agents in breast cancer patients. Polymorphisms in genes encoding Phase I – Cytochrome P450 (CYP450) and NAD(P)H dehydrogenase quinine (NQO1); phase II glutathioneStransferase (GST) and methylene tetra hydrofolate reductase (MTHFR); phase IIIp-glycoprotein (ABCB1) and solute transporter (SLC22A16) drug metabolizing enzymes can possibly predict clinical outcomes, and can be of prognostic significance in breast cancer patients. The aim of this study was to determine the role of genetic variations in drug metabolizing enzymes in predicting response and toxicity in breast cancer patients, using multi-analytical approaches.

Deciphering the Preference and Predicting the Viability of Circular Permutations in Proteins

Circular permutation (CP) refers to situations in which the termini of a protein are relocated to other positions in the structure. CP occurs naturally and has been artificially created to study protein function, stability and folding. Recently CP is increasingly applied to engineer enzyme structure and function, and to create bifunctional fusion proteins unachievable by tandem fusion. CP is a complicated and expensive technique. An intrinsic difficulty in its application lies in the fact that not every position in a protein is amenable for creating a viable permutant. To examine the preferences of CP and develop CP viability prediction methods, we carried out comprehensive analyses of the sequence, structural, and dynamical properties of known CP sites using a variety of statistics and simulation methods, such as the bootstrap aggregating, permutation test and molecular dynamics simulations. CP particularly favors Gly, Pro, Asp and Asn. Positions preferred by CP lie within coils, loops, turns, and at residues that are exposed to solvent, weakly hydrogen-bonded, environmentally unpacked, or flexible. Disfavored positions include Cys, bulky hydrophobic residues, and residues located within helices or near the protein's core. These results fostered the development of an effective viable CP site prediction system, which combined four machine learning methods, e.g., artificial neural networks, the support vector machine, a random forest, and a hierarchical feature integration procedure developed in this work. As assessed by using the hydrofolate reductase dataset as the independent evaluation dataset, this prediction system achieved an AUC of 0.9. Large-scale predictions have been performed for nine thousand representative protein structures; several new potential applications of CP were thus identified. Many unreported preferences of CP are revealed in this study. The developed system is the best CP viability prediction method currently available. This work will facilitate the application of CP in research and biotechnology.

Evaluation of C677T polymorphism of the methylenetetra hydrofolate reductase gene and its association with levels of serum homocysteine, folate, and vitamin B12 as maternal risk factors for Down syndrome

AIMS AND OBJECTIVE:Evaluation of C677T polymorphisms of the methylenetetra hydrofolate reductase (MTHFR) gene and its association with level of serum homocysteine, folate, and vitamin B12 as possible maternal risk factors for Down syndrome.DESIGN:This was a case–control study.MATERIAL AND METHODSFifty-two mothers (mean age 27.6 years) with babies having free trisomy 21 of North Indian ethnicity and 52 control nonlactating mothers (mean age 24.9 years) of same ethnicity attending services of genetic lab for bloodletting for other causes were enrolled after informed written consent. Fasting blood was collected and was used for determination of plasma homocysteine, vitamin B12, and folate (serum and RBC), and for PCR amplification of the MTHFR gene.RESULTS:The prevalence of MTHFR C677T polymorphism in north Indian mothers of babies with trisomy 21 Down syndrome was 15.38% compared to 5.88 % in controls. The difference between two groups was not statistically significant (P = 0.124). Low serum folate was demonstrated in 34.62% of cases vs. 11.54% in controls, which was significant (P = 0.005). Low RBC folate was found in 30.7% of cases versus 11.53% in controls, which was not significant (P = 0.059), when analyzed independently. But on multiple regression analysis the difference was statistically significant. Low serum vitamin B12 was found in 42.31% of cases versus 34.62% in controls, which was not significant (P = 0.118). The mean serum homocysteine in cases was 10.35 ± 0.68 while controls were 9.02 ± 0.535.CONCLUSION:Serum levels of folate were low in cases. The RBC folate levels were comparable in both groups. However the combined serum folate and RBC folate were low in cases compared to control groups. Homocysteine levels in our study were higher in Down syndrome mothers compared to controls; however high-serum level of Homocysteine had no association with MTHFR polymorphism. No association of serum vitamin B12 with MTHFR polymorphism in occurrence of Down syndrome births was found. Peri- or preconceptional folate supplementation may therefore lead to a decline in DS births, if supported by larger studies.

Relationship Between Genetic Polymorphisms of Methylenetetra hydrofolate Reductase (C677T, A1298C, and G1793A) as Risk Factors for Idiopathic Male Infertility

The human methylenetetrahydrofolate reductase (MTHFR) gene plays a crucial role in folate metabolism. Data regarding the influence of MTHFR gene polymorphisms on male fertility status are scarce and conflicting. We determined associations between 3 MTHFR gene polymorphisms (C677T, A1298C, and G1793A), serum folate, and total homocysteine (tHcy) levels, with male fertility status and semen parameters. MTHFR genotypes were determined using polymerase chain reaction restriction fragment length polymorphism (PCR-RLFP) technique and serum tHcy, folate, and vitamin B12 concentrations were measured in 164 men with idiopathic infertility and 328 healthy participants. There was a significant difference in genotype frequency distribution of MTHFR C677T polymorphism between infertile patients and controls (P = .004). The 677T allele carriers (TC or TT) had a significantly increased risk of infertility compared with the CC homozygotes (odds ratio [OR] 1.60, 95% confidence interval [CI] 1.21-2.75, and OR = 2.68, 95% CI = 1.84-3.44, respectively), in a logistic regression model after adjustment for confounding factors. Men with the 677T, 1298C, and 1793G alleles showed significantly higher serum tHcy and lower folate levels (all Ps < .01). We found a positive correlation between serum folate concentrations and sperm density (r = .74, P = .001), percentage of sperm with progressive motility (r = .68, P = .001), as well as percentage of sperm with normal morphology (r = .72, P = .001). MTHFR C677T polymorphism is associated with an increased risk of idiopathic male infertility. Further study on the biologic role that this polymorphism plays in the development of infertility may lead to better understanding of the etiology of impaired spermatogenesis.

Hypercoagulable State and Methylenetetra- hydrofolate Reductase (MTHFR) C677T Mutation in Patients with Beta-Thalassemia Major in Kuwait

Introduction: Patients with thalassemia major often present with a hypercoagulable state, the pathogenesis of which is still not understood. Materials and Methods: This study evaluates the risk factors for hypercoagulability in 50 β-thalassemia major patients and 50 healthy controls. Fasting total homocysteine, protein C (PC), protein S (PS), antithrombin (AT), activated protein C resistance (APCR) and lupus anticoagulant (LA) were assessed. MTHFR C677T mutation was determined. Results: Significant reductions in PC, PS and AT were noted in patients. Only 4% of the patients had hyperhomocysteinemia. Thirty-two percent of the patients were heterozygous and 4% were homozygous for MTHFR C677T mutation. Conclusion: The natural coagulation inhibitors PC, PS and AT were significantly reduced in patients with β-thalassemia major and were thus important risk factors for the hypercoagulable state, but hyperhomocysteinemia and MTHFR mutation do not seem to be significant risk factors for thromboembolic events.

Study of correlationship between congenital heart disease and 5, 10-methylenetetra hydrofolate reductase gene's polymorphism or folacin intakes

To investigate the correlationship between congenital heart disease and 5, 10-methylenetetra hydrofolate reductase (MTHFR)'s C677T or folacin intakes, and to study the interaction of them in the occurring of congenital heart disease. We used case-control study (case = 104, control = 208) method. Cases and controls were chosen by age, sex and other conditions. The MTHFR C677T genotype distribution was analyzed by using polymerase chain reaction restricted fragment length polymorphism (PCR-RFLP), and non-conditional and multi-conditional logistic regression analysis were also used to analyze the correlationship and interaction of the factors. In case group, the number of people in low folacin intake level was 38 (36.54%), which in control group was 21(10.10%). The intake level of folacin during pregnancy was related to congenital heart disease (chi(2) = 31.614, nu = 1, P < 0.0001). The value of OR was 1.417 with 95%CI 1.216 - 1.651, indicating that the low level of folacin intakes was a risk factor to the congenital heart disease. In case group, the number of TT genotype was 46 (44.24%), the number of CT genotype was 42 (40.38%), the number of CC genotype was 16 (15.38%). In control group, the number of TT genotype was 39 (18.75%), the number of CT genotype was 114 (54.81%), the number of CC genotype was 55 (26.44%). A significant genotype distribution difference was identified between case and control group (chi(2) = 23.13, nu = 2, P < 0.0001). Genotype MTHFR 677TT was a risk factor of congenital heart disease and the OR value was 3.437 (95%CI: 2.042 - 5.784). The interaction analysis suggested that the low level of folacin intakes and the MTHFR 677TT genotype had a positive adding effect in the occurring of congenital heart disease. After adjusted some factors such as the ages of parents, fetus age and sex, the effect values of interaction were 13.343 and 15.911 respectively, and the percentages of attributable interaction effects were 0.619 and 0.612. The percentages of effect values of interaction between pure factors were 0.649 and 0.637 and the population attributable risks were 25.26% and 27.82% according to the estimated exposure rate of population risk factors. The low level of folacin intakes during pregancy should be a risk factor to congenital heart disease and the MTHFR 677TT genotype be correlated to congenital heart disease. There is interaction between folacin intakes and the MTHFR 677TT genotype. Increasing the intakes of folacin among MTHFR 677TT genotype people might decrease the incidence rate of congenital heart disease.

The impact of genetic factors in the development of thrombosis in cancer patients

e22227 Background: Thromboembolism is frequent in patients with cancer. Although it is known that several acquired factors take place in this process, the role of the genetic factors is controversial. In this study we analysed the most common genetic polymorphisms which have a role in the development of thrombosis. Methods: Study population consists of 292 (158 with thrombosis and 134 without thrombosis) patients treated between 2004 and 2008. Thrombosis was diagnosed by clinically and radiological measures in any time during the course of disease. Factor V Leiden G1691A, Prothrombin G20210A, Methlene Tetra Hydrofolate Reductase (MTHFR) C677T and Plazminogen Activator Inhibitor (PAI-1) 4G/5G were analysed with PCR-RFLP (Polymerase Chain Reaction- Restriction Fragment Length Polymorphism). Results: In the study group 48 and 1 patient had heterozygot and homozygot polymorphism for Factor V Leiden G1691A respectively. In the control group 32 heterozygot and 1 homozygot polymorphism and there was no statistically significant differents between study and control group (p=0,462).For Protrombin G20210A 11 and 4 heterozygot polymorphism were observed in control and study group respectively. While there was 1 homozygot polymorphism in study group there was no in control group and the difference between two groups was not statistically significant (p=0,198). For MTHFR C677T, 48 and 24 heterozygot polymorphism, 15 and 12 homozygot polymorphism were observed in study and control group respectively. The difference between two groups were statistically significant (p=0,04). There was no statistically significant difference between two groups for PAI-1 4G/5G polymorphisms (p=0,362). In subgroup analyses, statistically significant difference was found only in MTHFR C677T polymorphism in patients with GI cancer and with and without thrombosis (p=0,028). Conclusions: The studies investigating the relationship between genetic factors and thrombosis revealed controversial results. However, we found no genetic factor relevant to thrombosis other than MTHFR C677T polymorphism. Further studies investigating genetic and acquired factors in the development of the thrombosis in detail are warranted for documenting clearly the role of genetic polymorphisms. No significant financial relationships to disclose.

Plasma Homocysteine and Gene Polymorphisms Associated with the Risk of Hyperlipidemia in Northern Chinese Subjects

To examine the relationship between occurrence of hyperlipidemia, plasma homocysteine and polymorphisms of methylenetetra hydrofolate reductase (MTHFR) gene and methionine synthase (MS) gene. A total of 192 hyperlipidemia patients were selected and divided into hypercholesterolemia group, hypertriglyceridemia group, and combined hyperlipidemia group. Another 208 normal individuals were selected as control. Total plasma homocysteine (tHcy) concentration was measured by high-performance liquid chromatography (HPLC). Lipid profiles were measured for all subjects. The polymorphisms of MTHFR gene C677T and MS gene A2756G were analyzed by PCR-RFLP. The tHcy concentration in the combined hyperlipidemia patients was significantly higher than that in the control (15.95 micromol/L vs 13.43 micromol/L, P < 0.05). The prevalence of hyperhomocysteinemia (HHcy) in the combined hyperlipidemia group was significantly higher than that in the control (42.2% vs. 23.0%, P = 0.015), with the odds ratio (OR) of 3.339 (95% CI: 1.260-8.849). The hyperlipidemia patients with HHcy had a higher concentration of total cholesterol (TC) than that in the normal tHcy patients (5.67 +/- 0.95 mmol/L vs. 5.47 +/- 0.92 mmol/L, P=0.034). There was no significant difference in genotype or allele frequencies of MTHFR C677T between the hyperlipidemic and control groups. The hyperlipidemia patients with MTHFR CT/TT genotype had a higher concentration of triglyceride (TG) than those with CC genotype (2.24 +/- 1.75 mmol/L vs 1.87 +/- 0.95 mmol/L, P < 0.05). Individuals with CT/TT genotype had a higher concentration of tHcy than those with 677CC genotype both in the hyperlipidemia group (12.61 +/- 1.24 micromol/L vs. 11.20 +/- 1.37 micromol/L, P < 0.05) and in the control group (14.04 +/- 1.48 micromol/L vs. 12.61 +/- 1.24 micromol/L, P < 0.05). The percentage of MS 2756 GG/AG genotype in the combined hyperlipidemia group was significantly higher than that in the control (26.7% vs. 13.0%, P = 0.012), with the OR of 3.121 (95% CI: 1.288-7.651). The hyperlipidemia patients with MS 2756AG/GG genotype had a higher concentration of TC (5.87 +/- 0.89 mmol/L vs. 5.46 +/- 0.93 mmol/L, P < 0.05) and LDL-C (3.29 +/- 0.81 mmol/L vs. 2.94 +/- 0.85 mmol/L, P < 0.05) than those with AA genotype. However, individuals with 2756AG/GG genotype showed no significant difference in tHcy among those with AA genotype. HHcy and MS A2756G mutation may be the risk factors for combined hyperlipidemia. Further study is needed to confirm the role of HHcy and MS A2756G mutation in the development of hyperlipidemia.

Investigation of the association between 677C&gt;T and 1298A&gt;C 5,10-methylenetetra- hydrofolate reductase gene polymorphisms and normal-tension glaucoma

Homozygous polymorphism of the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene and resultant hyperhomocysteinaemia have been established as an independent risk factor for vascular diseases. There are evidences that vascular abnormalities are involved in the pathogenesis and progression of normal-tension glaucoma (NTG). In the present study, we were to find out the associations between 677C>T and 1298A>C polymorphisms of the MTHFR gene and NTG. This was a retrospective, case-controlled study enrolling 78 NTG patients and 100 controls. DNA from peripheral blood lymphocytes was extracted and the genotypes of polymorphisms (677C>T and 1298A>C) in the MTHFR gene were determined using PCR followed by restriction enzyme digestion. The frequencies of the polymorphic genotypes in the patients with NTG and controls were compared. The frequencies of the polymorphisms of the MTHFR gene (677C>T and 1298A>C) in the NTG patients were not significantly different from those of controls. But the younger NTG patients (age at diagnosis < or = 45 years) showed significantly higher prevalence of 677C>T polymorphism than the older NTG patients (age at diagnosis > 45 years) (TT genotype, 38.9 vs 11.9%, P=0.006, OR=4.71, 95% CI=1.49-14.9) and than the younger control subgroup (TT genotype, 38.9 vs 6.1%, P=0.001, OR=9.86, 95% CI=2.23-42.4). The 677C>T polymorphism was significantly associated with NTG in the younger patients, while 1298A>C polymorphism was not. This suggests that 677C>T polymorphism of the MTHFR gene can be a genetic risk factor of NTG in Korean population.

Prevalence of the C677T Methylenetetra- hydrofolate Reductase Mutation in Thai Patients with Deep Vein Thrombosis

We investigated the prevalence of a genetic variation in the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene (C677T) using polymerase chain reaction techniques in a sample of 500 general Thai population and among 40 unselected Thai patients with an objectively confirmed history of deep vein thrombosis (DVT). The prevalence of the mutated homozygous and heterozygous C677T MTHFR genotype in the group of 500 healthy Thai population was 1.4 and 25.6%, respectively (allele frequency of 14.2%). Of the 40 patients studied, none were homozygotes and 15% were heterozygotes for the C677T MTHFR gene mutation (allele frequency of 7.5%). There was no significant difference in genotype frequency between patients and control groups (p = 0.09). Odds ratios for the probability of the C677T MTHFR gene mutation in the patient versus control group were 0.49 (95% CI 0.21–1.12). These data indicated that the C677T MTHF gene mutation was not associated with DVT in the Thai population. The lower frequency of the C677T MTHFR gene mutation in our Thai population compared with reports from other studies suggests a wide heterogeneity in the 677T MTHFR genotype frequencies of the different ethnic populations even among Asians.