ARMS-PCR Method Research Articles

Relationship between Functional miR-143/145 Cluster Variants and Susceptibility to Type 2 Diabetes Mellitus: A Preliminary Case-Control Study and Bioinformatics Analyses

ABSTRACT Purpose: To investigate the link between two variants (rs4705342 and rs4705343) in the promoter of the miR-143/145 cluster with Type 2 diabetes mellitus (T2DM) risk. Methods:A total of 1200 subjects were genotyped using the ARMS-PCR method. Results: The rs4705342 variant enhanced the risk of T2DM under codominant CC (OR = 3.24; 95% CI: 1.89–5.60), recessive TT+TC (OR = 3.02; 95% CI: 1.77–5.17), and dominant TC+CC (OR = 1.35; 95% CI: 1.08–1.71) genetic models. Individuals carrying the C allele of rs4705342 conferred a 1.43 fold increased risk of T2DM. As regards rs4705343, decreased risk of T2DM was observed under codominant TC (OR = 0.53; 95% CI: 0.42–0.67), over-dominant TT+CC (OR = 0.51; 95% CI: 0.40–0.64), and dominant TC+CC (OR = 0.59; 95% CI: 0.48–0.75) models. Haplotype analysis of the variants showed a 1.941-fold increased risk of T2DM regarding the C T combination. Significant associations were noticed between different haplotypes and lipid indices of T2DM patients. There were no notable changes in p-values after adjustment for BMI. Computational analysis revealed that miR143 and/or miR145 target important genes involved in glucose and lipid metabolism. Conclusions: Functional miR-143/145 variants might influence the risk of T2DM. Hence, clarifying the precise regulatory mechanisms of gene expression in the development of T2DM will significantly guide researchers to find a novel target for therapeutic intervention.

The efficacy of calcitriol treatment in non-alcoholic fatty liver patients with different genotypes of vitamin D receptor FokI polymorphism

BackgroundVitamin D deficiency is prevalent in patients with non-alcoholic fatty liver disease (NAFLD), but there are debates on the usefulness of vitamin D treatment. The interindividual variations in response may be due to different genetic backgrounds.The present study evaluated the efficacy of calcitriol treatment in NAFLD patients with regard to the vitamin D receptor (VDR) genotypes of FokI polymorphism.MethodsThe study was conducted on 128 NAFLD patients randomly divided into two groups and were subjected to intervention with 0.25 mcg calcitriol/day or placebo for 4 months, while anthropometric parameters, glycemic status, lipid profiles, inflammatory markers, liver enzymes, and fatty liver indices were measured. The ARMS-PCR method was used to genotype the VDR FokI polymorphism.ResultsCalcitriol treatments along with weight loss and diet recommendations decreased the liver enzymes (AST, ALT, and ALP, p < 0.001 for all) and fatty liver indices (HSI, p < 0.01 and APRI, p < 0.001), compared to the baseline. But when the calcitriol effects were compared to the placebo group, only ALP decrease remained significant (17.5 IU. P = 0.02). The prevalent FokI variants in our population were FF (53.1%) and Ff genotype (45.3%). No significant interaction of FokI variants to the calcitriol effects was found except for ALP. The decrease in the ALP activity was higher in calcitriol-received patients with the Ff genotype (p = 0.05).ConclusionsThe FF and Ff variants of VDR FokI polymorphism did not interact with the effects of calcitriol on fatty liver, but the ALP was more responsive in subjects with the Ff variant.IRCT registration numberIRCT2017053034222N1 Registration date: 2017-06-28 - Retrospectively registered, https://en.irct.ir/trial/26203

Association of BHMT (rs 3733890) gene polymorphism with biochemical markers of B12 deficiency in T2DM patients on metformin therapy

Long term administration of metformin interferes with the absorption of vitamin B12 resulting in vitamin B12 deficiency.Among the various genetic variants that are associated vitamin B12 deficiency, we analyzed BHMT gene polymorphism and their association with metformin induced vitamin B12 deficiency in T2DM patients.A cross sectional study was done with 300 participants. Methyl malonic acid, homocysteine and high sensitive C reactive protein were analysed by Mass Spectrometry, Chemiluminescent analyser and Immunoturbidimetric method respectively. Genetic variants were analysed by ARMS-PCR method, Data was analyzed with various statistical tools like ROC, Odds ratio and Likelihood ratio. Significant reduction in folic acid and vitamin B12 in metformin users was found. High sensitive C reactive protein, homocysteine and methyl malonic acid are significantly increased in patients with metformin induced B12 deficiency. ‘A’ allele in BHMT (A allele OR =2.1, AA genotype =2.8) showed risk of vitamin B12 deficiency in T2DM patients on metformin therapy. BHMT gene polymorphism had LR of 2.65 for folic acid (AG genotype), 2.73 for MMA (AA genotype) and 2.63 for Homocysteine (AA genotype).We found an association between single nucleotide polymorphism of BHMT and diagnosis of vitamin B12 deficiency status in metformin users. Folic acid, MMA and homocysteine had high specificity in concordance with BHMT (rs3733890) polymorphism in predicting vitamin B12 deficiency. Early screening of SNP of BHMT in T2DM patients on metformin therapy will help us to identify group of people who are prone for vitamin B12 deficiency.

Interlukine-17 Polymorphism in Patients with Inflammatory Bowel Disease ( IBD ) in Al-Anbar Province

Tetra amplification refractory mutation system polymerase chain reaction technique (tetra ARMS PCR)ARMS-PCR method was used to genotyping of IL-17A and IL-17F , The impact of IL-17A and IL-17Fgenotypes on the production of interleukins in IBD patients and control groups were determined andInterleukins gene polymorphisms role in IBD infection. significant deference at (p ? 0.05 ) for each rs763780in IL-17 F in patients especially Genotype TT,TC,CC were 31 (51.67%),19 (31.67%),10 (16.67%) \ andrs2275913 in IL-17 A in patients especially Genotype GG,GA,AA were 18 (30%),23 (38.33%),19 (31.67%).As well as genotyping of rs763780 in IL-17 F showed significant increase at a significant level of P?0.05in both TC and CC genotypes in the cases samples compared with the control samples. the odds ratio was2.8894 and 22.3333 for TC and CC genotypes respectively at confidence interval up to 7.8 and 397 forboth genotypes. T allele are more frequent in both cases and control and it have odds ratio as 5.021 Andgenotyping of rs2275913 in IL-17 A showed significant increase at a significant level of P?0.01 for the AAgenotyping in the cases samples compared with the control samples, while the GA genotyping did not showany significant differences, ), the odds ratio for GA genotypes was 2.3590 at confidence interval up to 5.8,while it was 8.4444 at confidence interval up to 32.9 for AA genotypes. G allele are more frequent in bothcases and control and it have odds ratio as 3.319 at confidence interval up to 6

Association of Novel Single Nucleotide Polymorphisms of Genes Involved in Cell Functions with Male Infertility: A Study of Male Cases in Northwest Iran.

Background:Infertility is a global health problem caused by various environmental and genetic factors. Male infertility accounts for 40–50% of all cases of infertility and approximately half of them are grouped as idiopathic with no definitive causes. Previous studies have suggested an association between some SNPs and infertility in men. In this study, an attempt was made to investigate the association of 7 different SNPs of 4 genes involved in common cell functions with male infertility.Methods:MTHFR rs1801131 (T>G), MTHFR rs2274976 (G>A), FASLG rs80358238 (A>G), FASLG rs12079514 (A>C), GSTM1 rs1192077068 (G>A), BRCA2 rs4987117 (C>T), and BRCA2 rs11571833 (A>T) were genotyped in 120 infertile men with idiopathic azoospermia or severe oligospermia and 120 proven fertile controls using ARMS-PCR methods. Next, 30% of SNPs were regenotyped to confirm the results. Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated using SPSS statistical software to evaluate the strength of association. The p<0.05 were considered statistically significant.Results:Statistical analysis revealed significant association between MTHFR rs-2274976 AA variant (OR: 10.00, CI: 3.203–31.225), FASLG rs12079514 AC variant (OR: 0.412, CI: 0.212–0.800), and BRCA2 rs11571833 TT variant OR: 6.233, CI: 3.211–12.101) with male infertility, but there was no significant difference between case and control groups in MTHFR rs1801131 (p= 0.111), GSTM1 rs1192077068 (p=0.272), BRCA2 rs4987117 (p=0.221), and FASLG rs80358238 (p=0.161).Conclusion:Our findings suggested that some novel polymorphisms including MTHFR rs2274976, FASLG rs12079514, and BRCA2 rs11571833 might be the possible predisposing risk factors for male infertility in cases with idiopathic azoospermia.

-2518A/G polymorphism of monocyte chemotactic protein 1 (MCP-1/CCL2) is associated with cutaneous mastocytosis.

IntroductionMastocytosis is a rare heterogeneous disease associated with pathological accumulation of mast cells (MCs) in one or more organs. The disease may be limited to the skin (cutaneous mastocytosis – CM), or present an internal organ involvement (systemic mastocytosis – SM). Pathophysiology of the disease is not well established. However altered proliferation, differentiation and chemotaxis of MC may play an essential role in the development of the disease. The monocyte chemotactic protein 1 (MCP-1/CCL2) may be one of the factors responsible for MCs migration to the skin and other organs.AimTo analyse the frequency of biallelic A/G polymorphisms at position -2518 in the promoter of the MCP-1 gene and compare the serum level of MCP-1 in patients with both forms of mastocytosis and the healthy control group.Material and methodsUsing ARMS-PCR methods we analysed -2518A/G polymorphisms in the promoter region of the MCP-1 gene in 127 mastocytosis patients (95 CM and 32 SM), and 160 healthy controls. Additionally, the MCP-1 serum level was detected with ELISA technique in 70 patients and 40 controls.ResultsWe have found that CM patients have more frequently the GG genotype of the MCP-1 gene (p = 0.01) in comparison to SM patients and controls. The GG genotype was more frequent in children than in adults (p = 0.02). The MCP-1 serum level was higher in SM patients than in CM patients and controls.ConclusionsResults of this study indicate that cutaneous mastocytosis could be associated with the -2518 A/G MPC-1 gene polymorphism.

TRAILR1 (rs20576) and GRIA3 (rs12557782) are not associated with interferon-β response in multiple sclerosis patients.

Multiple sclerosis (MS) is an autoimmune-type inflammatory disorder in human central nervous system. Recombinant interferon beta (IFN-β) decreases the number of relapses and postpones disability progression in MS. However, up to 50% of patients treated with interferon beta continue experiencing relapses and/or worsening disability. Single nucleotide polymorphisms in different genes have been known to show significant associations with response to IFN-β in MS patients. In the present work, we examined the potential role of TRAILR1 and GRIA3 genes polymorphisms on response to IFN-β therapy in Iranian MS patients. The DNA was extracted from blood samples by standard procedures from 73 patients diagnosed with Multiple Sclerosis that were either responded to IFN-β or did not. We carried out RFLP -PCR and tetra-primer ARMS-PCR methods to study of rs20576 and rs12557782, respectively. All results were analyzed using the SPSS software. TRAILR1 rs20576 genotype frequencies in responders and non-responders were similar (χ2 = 0.26, P = 0.87, Fisher, s Exact test). Our results showed that response to IFN-β has not association with sex (p = 0.73). Also, genotypic frequencies of GRIA3 rs12557782 had no significant differences between two groups of female population (χ2 = 3.75, p = 0.15). Furthermore, it had not been any statistical differences between responder and non-responder males (χ2 = 0.7, p = 0.4) related to the SNP. Our results analysis revealed no significant association between the studied SNPs (TRAILR1 rs20576 and GRIA3rs 12,557,782) and response to IFN-β in Iranian MS patients.

Evaluation of the correlation between serum lipid characteristics of obese subjects and ADIPOQ gene rs266729 polymorphism in Chaharmahal and Bakhtiari Province of Iran

Obesity is a multifactorial disorder that is influenced by various factors such as behavior, diet, environment, metabolic and genetic. This disease is the result of an imbalance between energy absorption and expenditure. Mutations in genes that are responsible for appetite control and metabolism are considered as the genetic component of obesity. Adiponectin protein is one of the most effective adipokines in regulating the body's energy homeostasis and fat storage, which is expressed by the ADIPOQ gene and secreted from white adipose tissue. The concentration of this protein in the blood decreases in obesity. In this study, the relationship between rs266729 polymorphism in the ADIPOQ gene with the level of biochemical parameters such as total cholesterol and triglyceride and HDL and LDL in the blood of obese people in Borujen (a city in Iran) was investigated. This study was performed on 100 people who referred to the Tamin Ejtemaee clinic due to obesity problems in Borujen. In this study, the ARMS-PCR method was used to determine the genotype of individuals. Based on the results of this study, no significant relationship was found between biochemical parameters including total cholesterol, triglyceride, and LDL with rs266729 polymorphism genotypes in ADIPOQ gene in obese subjects. We concluded that rs266729 polymorphism cannot be useful as an index parameter for predispose genotype for imbalance in total cholesterol, triglyceride, and LDL levels in a person.

Involvement of single nucleotide polymorphisms in acute lymphoblastic leukemia susceptibility

Acute lymphoblastic leukemia (ALL) is an overgrowth of lymphoid progenitor cells and accounts for the highest common type of childhood cancer. Early detection of ALL could significantly improve the treatment and thereby survival of the patients. Single Nucleotide Polymorphisms (SNPs) have been emerging as effective diagnostic biomarkers in cancer. However, little is known about the association between the SNPs and the risk of ALL. In this study, we genotyped ALL patients and controls for eight SNPs, using the Tetra-primer ARMS PCR method. The results showed there were significant associations between the rs1333048, rs2070672, rs2279744, rs3733890, rs11549465 and the risk of ALL cancer. The findings may have diagnostic implications for early detection of ALL.

ESR1 gene variants, haplotypes and diplotypes may influence the risk of breast cancer and mammographic density.

Breast cancer as the most common cancer worldwide is influenced by genetic and physiological factors. Based on some evidence indicating the role of estrogen receptor 1 gene (ESR1) in breast cancer development, in this study, the association of three common variations in ESR1 gene with breast cancer and density in an Iranian population was evaluated. In a case-control study, 400 blood samples were collected for DNA extraction and genotyping. Breast density was assessed using mammography. ESR1 rs6915267 (G/A), rs2077647 (C/T) and rs1801132 (C/G) were genotyped using ARMS-PCR method. PHASE program was used to estimate the haplotypes frequencies. Our data analysis showed rs6915267 GA genotype in the heterozygous (GA) as well as co-dominant models was associated with lower mammographic density. None of the three variations were associated with the breast cancer risk. Haplotype analysis indicated G-T-C haplotype of rs6915267, rs2077647 and rs1801132 [OR = 0.54, 95% CI (0.31-0.92), p = 0.025] and G-T/G-T diplotype of rs6915267-rs2077647 [OR = 0.38, 95% CI (0.17-0.86), p = 0.019] were associated with a decreased risk of breast cancer. ESR1 may affect density of the breast and its haplotypes may modulate breast cancer risk.

LRP8 (rs5177) and CEP85L (rs11756438) are contributed to schizophrenia susceptibility in Iranian population

Introduction Schizophrenia is recognized as one of the most important mental illnesses of the last century. Many genetic and environmental factors are involved in this condition. Recently, the genome-wide association study identified two significant genes LRP8 and CEP85L associated with psychiatric disorders. LRP8 (low-density lipoprotein receptor-related protein 8) acts as a cytoplasmic receptor for Reelin. Many studies have revealed that LRP8 was significantly related to schizophrenia and bipolar disorder in Chinese population. CEP85L standing for 'centrosomal protein 85 kDa-like' is another gene, which has been reportedly associated with BPD. Methods We performed a case-control study to analyze the association between rs5177 single-nucleotide polymorphism in the LRP8 gene plus the single-nucleotide polymorphism rs11756438 in the CEP85L gene and schizophrenia in the Iranian population. The genotype for rs5177 was determined by ARMS PCR method, while for rs11756438 genotype, it was determined by PCR-RFLP method after which statistical analysis was performed for each polymorphism. In rs5177, the CC genotype was susceptible to the disease while G allele was associated with disease protection. Results and Conclusion In rs11756438, the AA genotype was associated with disease susceptibility, while allele A did not have a significant association with the disease.

IL-18 variant increases risk of enhanced HBV DNA replication in chronic hepatitis

BackgroundThe outcome ofhepatitis B (HBV) infection is influenced by immune responses and host genetics. Interleukin-18 (IL-18) is a determinant factor in controlling the balance of Th1/Th2 during antiviral response.Weexamine therole of two functional polymorphisms -607A/C and-137A/C inIL-18 gene with risk of chronic HBV infection. Methods and ResultsGenomic DNA isolates were obtained from 200 seropositive cases stratified according to their HBV DNA loads, and 200 blood donorsas a control population. Genotypes of the two polymorphisms were identified by ARMS-PCR method. The -607A allele, the-607AA and -607AC genotypes were associated with increased risk to develop chronic HBV infection (1.98, 5.11 and 3.5-fold risks, respectively). By contrast, the -137C minor allele and CG genotype had protected effects against chronic HBV infection.We found that -607A allele, -607AA and -607AC genotypes were significantly more frequent in patient’s group with high HBV DNA levels compared to patient group with low HBV DNA level. Additionally, they were associated with increased 1.72, 6.04 and 3.28-fold risk of high HBV DNA replication.Patients carrying “-607A/-137 C” or “-607A/−137 G” haplotypes presented a high risk to develop chronic HBV infection (OR = 3.27; OR = 4.32, respectively). ConclusionsTaken together, our data suggest that theIL-18 -607A/C functional polymorphism was associated with susceptibility to enhanced replicative form of HBV DNA in chronic infection.

Relationship Between ZAP-70 Gene (rs104893674 "A/C") Polymorphism With Risk for Acute Myeloid Leukemia

Background and Aim: In acute myeloid leukemia, a large number of immature cells develop, which can related to some single nucleotide polymorphisms presence in positions of genes that encodes enzymes involved in cell activation and evolution signaling pathways. In this study, the association of rs104893674 (A / C) polymorphism with the risk of Acute Myeloid Leukemia (AML) in samples obtained from Fars and Isfahan Province hospitals was investigated. Methods &amp; Materials: In the present case-control study conducted at Islamic Azad University of Kazerun in 2019, 94 AML patients and 99 age and sex-matched healthy individuals were enrolled. The rs104893674 (A / C) polymorphism was determined by Tetra Primer ARMS PCR method. Data were analyzed by SPSS (version23) software using Chi-square statistical test. Ethical Considerations: This study with research ethics code IR.IAU.KAU.REC.1398.051 has been approved by Research Ethics Committee of Islamic Azad University of Kazerun. Results: The results of this study showed a significant, allele and genotype-specific Association between rs104893674 (A / C) polymorphism with risk of AML. Thus, there are more likely to develop AML in AC genotype, individuals with A allele at this polymorphic site (P=0.000). Conclusion: The association of acute myeloid leukemia with the genetic polymorphism of the ZAP-70 protein can be considered as an option for prognosis of this complication in susceptible individuals.

SNPs in the 3'-untranslated region of SLC30A8 confer risk of type 2 diabetes mellitus in a south-east Iranian population: Evidences from case-control and bioinformatics studies.

Type 2 diabetes mellitus (T2DM) is a heterogenic disease with increasing incidence. The SLC30A8 gene encodes an islet zinc transporter (ZnT8), and its variants have been associated with glucose and pro-insulin levels. This study was aimed to examine the effects of a missense variant (rs13266634 C/T), and two 3'UTR variants (rs2466294 C/G and rs2466293 T/C) in SLC30A8 gene on T2DM risk in a south-east Iranian population. In this experiment, 450 patients diagnosed with T2DM and 453 healthy subjects from the same geographic area were enrolled. Genotypes were amplified using the ARMS-PCR method. In silico analyses were performed to determine the effects of the variants on the local structure of mRNA, splicing patterns, and potential miRNA-gene interactions as well. Significant differences were noticed between cases and controls regarding the genotypic and allelic distribution of the studied variants. As regards rs2466293 and rs2466294 variants, enhanced risk of T2DM was found under allelic, dominant, recessive, and codominant models (OR > 1). Besides, different genetic models of rs13266634 were associated with decreased risk of T2DM (OR < 1). Bioinformatics analyses indicated that the rs2466293 variant might influence the binding of some miRNAs, while the G-allele of rs2466294 decreased the stability of SLC30A8-mRNA. In our population, both SNPs in the 3'-untranslated region of SLC30A8 increased the risk of T2DM, while the rs13266634 variant showed a protective association against T2DM susceptibility. Investigating the effects of other variants in this gene or other ZnTs can further indicate such associations in subjects from the same ethnicity.

The association of methylene tetrahydrofolate reductase (MTHFR) A1298C gene polymorphism, homocysteine, vitamin B12, and folate with coronary artery disease (CAD) in the north of Iran

Abstract Background We pursued to find out the possible association of Methylene tetrahydrofolate reductase (MTHFR) A1298C gene polymorphism, blood homocysteine, vitamin B12, and folate with Coronary artery disease (CAD) in the study population in Guilan, north of Iran. Material and Methods Ninety patients with CAD and 76 healthy controls were evaluated. MTHFR A1298C polymorphism and its genotype frequency, the plasma level of homocysteine, vitamin B12 and folate were evaluated by using ARMS-PCR, ELISA, and Chemiluminescence methods, respectively. Results The frequency of genotypes, A, AC and CC in CAD were 40, 35.6, 24.4%, respectively which was significantly different (p=0.016) from the control group that were 26.3, 57.9 and 15.8%, respectively. The serum level of vitamin B12 and folate in genotype A1298C were not statistically significant between two groups (p&gt;0.05), however, the plasma homocysteine in patients with CAD was remarkably higher than the control group (p&lt;0.001). Additionally, in CAD patients the plasma level of homocysteine in the AC genotype was significantly higher than the control subjects (p=0.005). Conclusion It is thus concluded that MTHFR A1298C gene polymorphism is associated with CAD. It seems that the AC genotype of MTHFR A1298C polymorphism might have a protective effect on CAD.

Association of rs12487066, rs12044852, rs10735781, rs3135388, rs6897932, rs1321172, rs10492972, and rs9657904 Polymorphisms with Multiple Sclerosis in Iranian Population.

ObjectivesMultiple sclerosis (MS) is a chronic disease of the central nervous system. The pathogenesis of MS is best described by a multifactorial model incorporating interactions between genetic and environmental factors with the role of genetic factors increasingly taken into account. The main goal of this study was to investigate the associations of rs12487066, rs12044852, rs10735781, rs3135388, rs6897932, rs1321172, rs10492972, and rs9657904 polymorphisms with MS in the Iranian population.MethodsA total of 83 patients with MS (82.0% female and 18.0% male; mean age = 35.2±8.6 years) and 100 physically and mentally healthy subjects (81.0% female and 19.0% male; mean age = 40.4±6.4 years) were selected using convenient sampling. A 5 mL blood sample was taken from each case and control patient. We used the tetra-primer ARMS-PCR method to genotype the desired polymorphisms. The associations between polymorphisms and the disease were studied based on codominant, dominant, recessive, and overdominant models.ResultsThe rs10735781 polymorphism was codominantly (p = 0.029), overdominantly (p = 0.008), and dominantly (p = 0.009) associated with the disease. The rs6897932 was also found to be codominantly (p = 0.012), dominantly (p = 0.019), and recessively (p = 0.011) associated with the disease.ConclusionsWe found an association between the rs10735781 and rs6897932 polymorphisms on the EVI5 and IL7RA genes, respectively, with increased MS in the Iranian population. Therefore, single nucleotide polymorphisms in the EVI5 and IL7RA genes can be considered a prognostic marker of MS.

Association between PPARγrs1801282 polymorphism with diabetic nephropathy and type-2 diabetes mellitus susceptibility in south India and a meta-analysis

BackgroundDiabetic Nephropathy (DN) is a major complication of Type 2 Diabetes Mellitus (T2DM) with high morbidity rates worldwide. ObjectiveTo determine the association of PPARγ rs1801282 polymorphism in T2DM and DN in south Indian population. MethodsWe have conducted a case–control study to test the association of rs1801282 polymorphism with T2DM and DN in 424 subjects (DN=128; T2DM=148 and controls=148) belonging to the south Indian population using ARMS-PCR and Sanger sequencing method. Further, a meta-analysis was performed for rs1801282 polymorphism from the published literature retrieved from various electronic databases to determine the susceptibility among T2DM and DN across various ethnic populations under five genetic models. ResultsThe genotyping of rs1801282 polymorphism showed significant (p-value<0.05) association with DN and T2DM compared to controls. In the meta-analysis, no significant association (p-value>0.05) was noticed for rs1801282 with DN vs. controls in homozygote, heterozygote, allelic, recessive and dominant genetic models. However, a significant association was observed between rs1801282 SNP and T2DM under heterozygote (Jj vs JJ) genetic model with OR=0.56, (95%CI [0.43–0.74]), p≤0.0001 of Asian and Caucasian populations. ConclusionOverall analysis suggests that the rs1801282 polymorphism might be associated with DN and T2DM. More case–control studies on the PPARγ gene with a larger sample size including all the confounding factors are required to corroborate the findings from this meta-analysis.

Association between PPARγrs1801282 polymorphism with diabetic nephropathy and type-2 diabetes mellitus susceptibility in south India and a meta-analysis

BackgroundDiabetic Nephropathy (DN) is a major complication of Type 2 Diabetes Mellitus (T2DM) with high morbidity rates worldwide. ObjectiveTo determine the association of PPARγ rs1801282 polymorphism in T2DM and DN in south Indian population. MethodsWe have conducted a case–control study to test the association of rs1801282 polymorphism with T2DM and DN in 424 subjects (DN=128; T2DM=148 and controls=148) belonging to the south Indian population using ARMS-PCR and Sanger sequencing method. Further, a meta-analysis was performed for rs1801282 polymorphism from the published literature retrieved from various electronic databases to determine the susceptibility among T2DM and DN across various ethnic populations under five genetic models. ResultsThe genotyping of rs1801282 polymorphism showed significant (p-value<0.05) association with DN and T2DM compared to controls. In the meta-analysis, no significant association (p-value>0.05) was noticed for rs1801282 with DN vs. controls in homozygote, heterozygote, allelic, recessive and dominant genetic models. However, a significant association was observed between rs1801282 SNP and T2DM under heterozygote (Jj vs JJ) genetic model with OR=0.56, (95%CI [0.43–0.74]), p≤0.0001 of Asian and Caucasian populations. ConclusionOverall analysis suggests that the rs1801282 polymorphism might be associated with DN and T2DM. More case–control studies on the PPARγ gene with a larger sample size including all the confounding factors are required to corroborate the findings from this meta-analysis.

Kidd Blood Group Genotyping for Thalassemia Patient in Iran.

We aimed to determine the JK genotype in thalassemia patients from Iran using different molecular methods to compare with phenotyping results. We also aimed to standardize for the first time, the Tetra-Primer ARMS PCR method for JK genotyping. The serology method cannot correctly determine the phenotype of blood group antigens in patients with multiple blood transfusions. Peripheral blood samples were taken from two hundred alloimmunized thalassemic patients in Tehran Adult Thalassemic Clinic. The samples were tested phenotypically by routine serological methods. After DNA Extraction, SSP-PCR was performed. DNA sequencing and PCR-RFLP were used to confirm the SSP-PCR results. Discrepancies were found between the phenotype and genotype in 32 out of 200 cases. In 16 cases phenotype was determined as Jk (a + b +) but genotype was JK*A/JK*A, in 14 cases phenotype was Jk (a + b +) while the genotype showed JK*B/JK*B, 1 case had been phenotyped as Jk (a + b-) but it was genotyped as JK*A/JK*B and 1 case had been phenotyped as Jk (a-b +) but it was genotyped as JK*A/JK*B. Serological results for a few samples could not be confirmed because of mix-field agglutination. The genotyping however verified the presence of Kidd alleles. Molecular methods are a valuable tool to predict blood group phenotypes in multi-transfused patients in order to select RBC units for a perfect matching improving blood transfusion and preventing alloimmunization. Also Tetra-Primer ARMS PCR is simple and cost effective methods that could be alternative by conventional Molecular methods.

EGFR Gene Mutation and Methodological Evaluation in 399 Patients with Non-small Cell Lung Cancer.

The purpose of the present study was to study the characteristics of epidemic growth factor receptor (EGFR) gene distribution in patients with non-small cell lung cancer (NSCLC), and to detect the mutation rate of EGFR gene by Sanger sequencing and amplification refractory mutation system (ARMS)-PCR. Paraffin-embedded sections of NSCLC tissues from 399 NSCLC patients diagnosed in Renmin Hospital of Wuhan University were collected, 103 of them were detected for exons 18-21 mutation of EGFR by Sanger sequencing method, 296 cases were detected for exons 18-21 mutation by ARMS-PCR method. DNA extraction of both groups was performed with Qiagen QLAamp DNA FFPE Tissue KIT. Comparisons of detection rates between the two methods were conducted by row X list chi-square test. The total mutation rate of EGFR gene detected by Sanger sequencing was 21.4%, exons 18-21 and combined mutation rates were 1.0%, 9.7%, 1.0%, 7.8% and 2.0%, respectively. And the proportions were 4.7%, 45.2%, 4.7%, 36.3% and 9.4% respectively. The total mutation rate detected by ARMS-PCR was 51.4%, exons 18-21 and combined mutation rates were 2.7%, 27%, 1.7%, 18.2% and 1.7%, respectively. The proportions were 5.3%, 52.6%, 3.3%, 35.5% and 3.3% respectively. Further analysis of mutation rate showed that there was significant difference between the two methods in detecting total mutation of EGFR gene (P<0.001). There were significant differences in mutation detection rates of exons 19 and 21 (P<0.001, P<0.05), but there were no significant differences in other exons. And there was no significant difference in mutation detection rates between the two methods. The mutation rate of EGFR gene in NSCLC patients was 50%. And exon 19 deletion was the most common mutation type, followed by exon 21 mutation. Compared with Sanger sequencing method, ARMS method is more sensitive with significant advantages in detecting exon 19 deletions and exon 21 mutations, which can be widely used in clinical detection of EGFR gene mutations. The results of this study will further guide patients with advanced NSCLC to select TKI targeted drugs, and provide clinical diagnostic basis for targeted therapy of NSCLC patients.