Refractory Mutation System-polymerase Chain Research Articles

Association between interleukin-2 (rs2069762) gene polymorphism and FVIII inhibitor development in Indonesian patients with severe hemophilia A

BACKGROUND Factor VIII (FVIII) inhibitors in hemophilia A (HA) patients render FVIII replacement therapy ineffective. Although its development cause is unclear, it has been classified into therapeutic and genetic-related etiologies. Single nucleotide polymorphisms (SNPs) in several cytokine genes, including interleukin (IL)-2, could increase the risk of FVIII inhibitor development. This study aimed to evaluate the association between IL-2 (rs2069762) gene SNP and FVIII inhibitor development in Indonesian patients with severe HA.
 METHODS The IL-2 (rs2069762) gene SNP was examined in 119 HA patients. The presence of FVIII inhibitors was quantified using an enzyme-linked immunosorbent assay, with a titer of <0.28 ng/ml considered negative. Patients were divided into two groups: 59 with FVIII inhibitors (positive group) and 60 without inhibitors (negative group). The genotype of the subjects was determined using peripheral blood mononuclear cells and tetra-primer amplification refractory mutation system-polymerase chain reaction.
 RESULTS There was no association between IL-2 (rs2069762) gene polymorphism and FVIII inhibitor development on genotypes (p = 0.138) and allele frequencies (p = 0.780).
 CONCLUSIONS IL-2 (rs2069762) gene polymorphism is not a risk factor in the development of FVIII inhibitors in Indonesian patients with severe HA. Thus, further polymorphism studies in other cytokine genes are required to gain a comprehensive understanding of the FVIII inhibitor development.

Comparison of Allele Frequency of Uromodulin Gene rs13333226 and rs13333144 in a Sample of Iraqi Patients on Dialysis

Chronic kidney disease (CKD) is a major public health concern around the world. UMOD gene variants are linked to a higher incidence of hypertension and CKD in the general population. This study aimed to investigate the role of uromodulin rs13333226 and rs13333144 genes association with chronic kidney disease.The study samples were divided into two groups. The first group included 100patient samples and 70 chosen among them were under the dialysis and had kidney failure aged between 18-88 years old. The second group included 30 samples from healthy individuals who were used as a control. One of the ways used to identify the genotype is the tetra-primers amplification refractory mutation system–polymerase chain reaction (ARMS–PCR). Regarding the results of SNP (rs13333226 and rs13333144), the genotypes GG (OR=150.3), and AA (OR=0.01) for rs13333226. The genotypes GG (OR=0.02) and TT(OR=140.4) for rs13333144 when comparing between patients and control (P≤ 0.01), they were observed when the odds ratio is less than 1 means they are preventive factors, while OR more than 1 means the risk increase (P≤ 0.01). It was concluded that there was an association between SNP (rs13333226 and rs13333144) and uromodulin in kidney diseases. Where genotype GG) rs13333226) and TT (rs13333144) represents a risk factor and AA rs13333226 and GG rs13333144 represents a protective factor.

Study the Association of Uromodulin Gene rs13332878 with Chronic Kidney Disease

Uromodulin is the most abundant protein ordinary excreted in urine which could be used as a biomarker to diagnose kidney diseases. However, evidence suggests that it regulates salt transport, protects against urinary tract infection and kidney stones, and has a role in kidney damage and innate immunity. This study aimed to understand the association of uromodulin gene rs13332878 with chronic kidney disease. More than 100 people were selected for the study and the samples collected from the under study subjects were divided into two groups. 70 chosen subjects were under the dialysis with kidney failure, and aged between 18-88 years. The second group included 30 samples from healthy individuals, used as control. One of the ways used to identify the genotype was the tetra-primers amplification refractory mutation system–polymerase chain reaction (ARMS–PCR). Regarding the results of SNP (rs13332878), there were three genotypes: GG, CG and CC. Genotype GG was shown in the size 288bp, genotype CG was shown in the size 288+577bp, whereas genotype CC was shown in the size 577bp. The results of frequency genotypes that appeared for GG, GC and CC were 74.29%, 17.14% and 8.57% respectively in patients. Whereas in the control, the frequency of genotypes was GG 3.33%, GC 16.67% and CC 80%. It was observed that less than 1 odds ratio meant that the genotypes were to be considered as a preventive factor. While more than 1 OR meant that the risk had increased (P-value 0.01). No significant differences for the genotypes GC were found when comparing patients and the control groups (P> 0.01). So, we can conclude that there was an association between SNP (rs13332878) of uromodulin with kidney disease and genotype GG should be considered a risk factor while CC genotype represents a preventive factor.

Polymorphism in SNP rs972283 of the KLF14 gene and genetic disposition to peptic ulcer

Introduction and Aim: Kruppel Like Factor 14 (KLF14) gene plays an important role in metabolic illnesses and is also involved in the regulation of many other biological processes. This study's objective was to determine whether or not the KLF14 single-nucleotide-polymorphism (SNP) known as rs972283 was linked to an increased risk of peptic ulcer disease in the population that was being investigated. Materials and Methods: Participants in this study included 71 people who had been diagnosed with peptic ulcers and 50 people who were considered to be healthy controls. In order to genotype the KLF14 SNP rs972283, an amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) was carried out, and the PCR results were then sequenced. Results: Genotypes (GG, AG, and AA) were significantly different in people who had peptic ulcers compared to those who were in the control group (chi-square=7.703, 5.317 and 4.294) respectively. AG and AA genotypes for KLF14 gene were correlated with a high risk of peptic ulcer (P 0.05) (Odds Ratio (O.R.) =6.343 and 2.441) respectively. Patients with peptic ulcer had a significantly greater incidence of the allele A gene (30.3%), whereas healthy people had a much higher incidence of the G allele (86%). Patients who carried the AG genotype and had a chronic H. pylori infection were found to have a highly significant correlation with one another (P 0.01, O.R. =1.218). Similarly, there was a higher frequency of the G allele (84.6%), in people who had peptic ulcers, but there was a higher frequency of the A allele (39%), in cases of chronic infection. Conclusion: According to the findings of this research, a variant in the KLF14 gene called rs972283 is linked to an increased risk of peptic ulcer illness.

Tetra-ARMS-PCR assay development for genotyping of AGT rs699 T/C polymorphism, its comparison with PCR-RFLP and application in a case-control association study of cardiovascular disease patients

Genetic variations in the AGT gene play a significant role in controlling the plasma concentration of angiotensinogen (precursor protein of bioactive octapeptide angiotensin II) and the efficacy of antihypertensive drugs. In the current study, Tetra–Amplification Refractory Mutation System–Polymerase Chain Reaction (T-ARMS-PCR) was developed for genotyping of AGT rs699 T/C polymorphism and validated through Sanger DNA sequencing. Its efficiency was also tested using 474 human DNA samples [control, n = 181; cardiovascular disease (CVD) patients, n = 293]. Results showed that T-ARMS-PCR is superior to the commonly used PCR-Restriction Fragment Length Polymorphism (PCR-RFLP). Statistical analysis revealed that the AGT rs699 CC genotype is more prevalent in the CVD patient group (37% vs. 28%) and AGT rs699 C allele and CC genotype increased the risk of CVD by 1.4 and 1.9 fold, respectively. In summary, T-ARMS-PCR is the most suitable approach for quick and efficient genotyping of AGT rs699 T/C polymorphism in a large population in resource-limited countries, Furthermore, AGT rs699 T/C polymorphism is associated with the risk of CVD in the Punjabi Pakistani population.

Association of GSTTI, M1 and Polymorphism in GSTPI with Chronic Periodontal Disease in a Pakistani Population.

Chronic periodontal disease (CP) is a multifactorial infectious and inflammatory disease that occurs due to the challenge between the immune response of the host and specific periodontal bacteria, and that can lead to tooth loss due to damage inflicted to the supporting tissue. The current study investigates the genotypes of the GSTM1 and GSTT1 genes, along with the allelic frequency of the single nucleotide polymorphism [SNP; rs1695] in the GSTP1 gene and correlates them individually or in various combinations with the incidence of CP. A total of 203 clinically confirmed CP patients and 201 control subjects were enrolled from Multan and Dera Ghazi Khan Districts in Pakistan from April to July 2022. Multiplex Polymerase Chain Reaction (PCR) and tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR) approaches were applied to determine the genotypes of the studied GSTs. The association of rs1695 in GSTP1 with CP was studied both individually and in various combinations with GSTM1 and T1. The absence of GSTM1, the presence of GSTT1 and the presence of the mutant allele (G) at rs1695 in GSTP1 were found to be significantly associated with CP. Patients aged between 10 and 30 years were more affected by CP. Our results indicate that the genotypes of the analyzed GSTs affect the levels of protection from oxidative stress and may therefore influence the disease progression in CP.

Association of single nucleotide polymorphisms in heat stress protein 70 (HSP70) and 90 (HSP90) with the susceptibility of Pakistani sheep breeds to hemoparasitic infections

The present study was designed to report the genotypic and allelic frequency of single nucleotide polymorphism (SNP) at 222 G > A in HSP70 and at ex6-7390T22G in the HSP90 gene of 204 sheep (Baluchi = 11, Kajli = 29, Latti = 06 and Mundri = 158) enrolled from District Rajanpur in Punjab and to report the susceptibility of these sheep to the blood-borne parasitic infection. The tetra-primer amplification refractory mutation system–polymerase chain reaction (T-ARMS–PCR) approach revealed a significant variation (p < 0.001) in the genotype frequency of four enrolled sheep breeds at SNP 222 G > A in the HSP70 gene while the allelic frequency remained unaffected (p = 0.08). In all sheep breeds, GG (wild) genotype was most common. T-ARMS–PCR analysis revealed a similar trend for ex6-7390T22G in the HSP90 gene and it was observed that sheep had significantly higher wild-type (GG) (p < 0.05) at the studied SNPs. Studied epidemiological factors (sex and sampling sites) were not found associated with both SNPs. Chi-square test revealed that no specific genotype and allelic frequency at 222 G > A in HSP70 and at ex6-7390T22G in the HSP90 gene of the enrolled sheep breed was associated with the susceptibility to blood-borne parasitic infection (p > 0.05). In conclusion, we are reporting that Pakistan is blessed to have majority of sheep, from all breeds, having wild genotype at analyzed SNPs in heat stress genes. We highly recommend the genotypic screening of sheep before their selection as breeders to reduce the possibility of having sheep with polymorphic genotypes at 222 G > A in HSP70 and at 7390T22G in HSP90 genes that will improve the profitability and sustainability of animal production systems in Pakistan.

Identification of a Missense Mutation in GJA8 Gene in an Iranian Family with Autosomal Dominant Congenital Cataract.

To identify the causative mutations of autosomal dominant (AD) congenital cataracts in a large Iranian family. The complete and accurate family history and clinical information of participants were collected. A total of 51 family members, including 22 affected and 29 unaffected individuals, were recruited in this study. We performed whole exome sequencing to reveal pathogenic mutation. We used amplification refractory mutation system polymerase chain reaction and Sanger sequencing techniques to confirm segregation in patients and also to rule it out in the healthy participants. A known missense mutation, c.827C>T (S276F), in GJA8 was identified. This mutation was confirmed in all patients. Neither all healthy family members nor 100 healthy individuals who served as controls from general population had this mutation. The missense mutation c. 827C>T in the GJA8 gene is associated with AD congenital lamellar cataract with complete penetrance in a six-generation Iranian family.

Hyperfunction variant rs708035 of interleukin 1 receptor-associated kinases 2 gene involved in the predisposition of leprosy infection.

Mycobacterium leprae (slow-growing bacteria) is the etiological agent for leprosy infection, which is a chronic granulomatous disease. Symptoms initiate with the loss of sensation in the affected areas, which can lead to severe injuries, cuts and burns. IRAK2 (interleukin-1 receptor-associated kinases 2) is reported to function in the regulation of the NFκB pathway. The frequency of the IRAK2 polymorphism (rs708035) was unknown in the Pakistani population. Therefore, the study was designed to identify the role of the rs708035 SNP (single nucleotide polymorphism) in susceptibility to leprosy. The case-control study was designed, and participants were selected by Ridley-Jopling Classification. Blood samples from healthy individuals and patients were collected after ethical approval. Genomic DNA was extracted for the amplification of selected polymorphisms by tetra-primer amplification refractory mutation system polymerase chain reaction. The desired products were observed via agarose gel (2.5%) electrophoresis followed by data analysis using bioinformatics tools (SNP Stats and SHEsis) and statistical tests (odds ratio, OR, and chi square). The study revealed that the mutant genotype (TT) was found to be frequent among cases (22.80%) in comparison with the controls (1.66%). The SNP rs708035 was significantly associated with the progression of leprosy (χ2 = 17.62, p < 0.0001). The targeted SNP significantly increases the risk of leprosy 2.3 times (OR =2.3119, 95% CI1.2729-4.1989, p< 0.01). The genetic model also confirms the significant association of the A/T genotype with leprosy in the over-dominant model (OR =2.83, 95% CI1.16-6.89, p< 0.001). The study revealed a significant association of the targeted SNP with leprosy and provided baseline data regarding the association of rs708035. The current research could be utilized for the preparation of biomarkers by considering a larger sample size. The patients suffering from leprosy faced various comorbidities, including hypertension and diabetes. The study reports for the first time a significant association of interleukin 1 receptor associated kinases 2 (IRAK2) single nucleotide polymorphism (SNP) rs708035 among the Pakistani population (Karachi). The current study provides baseline data to develop diagnostic biomarkers for early detection of leprosy.

Identification of a Potential SNP Related to the Expression of Immune Genes and Its Possible Application to Selection of WSSV-Resistant Pacific White Shrimp (&lt;I&gt;Litopenaeus vannamei&lt;/I&gt;)

Highlight Research A molecular marker for shrimp selection in disease resistance. Single nucleotide polymorphisms (SNP) in the ALF gene strongly correlate with shrimp resistance to WSSV infection. Higher ALF gene expression in survivor shrimp. SNP as molecular marker inherited in the first generation (G1) shrimp. ARMS-PCR method successfully detect SNP in the shrimp ALF gene. Abstract The Pacific white shrimp (Litopenaeus vannamei) is Indonesia’s main export commodity, but its production is constrained by the white spot syndrome virus (WSSV). Selective breeding of disease-resistant broodstock based on single nucleotide polymorphism (SNP) in the anti-lipopolysaccharide factor (ALF) gene is an alternative strategy for solving the disease problem. This study aimed to detect the SNP g.455 A&gt;G in the anti-lipopolysaccharide factor (ALF) shrimp gene, evaluate the correlation of SNP with WSSV-resistance trait, analyze the expression level of immunity genes and genotype frequencies of the WSSV-resistance population shrimp and analyze the SNP inheritance in the first generation of selected shrimp. A total of 120 individuals from 4 families were used to detect the SNP marker using tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). The correlation of the SNP marker with survival rate (SR) was analyzed using a general linear model (GLM) between genotype frequencies and SR. Genotypic similarities between broodstock and pedigree were analyzed using Chi-square. SNP g.455 A&gt;G was successfully detected using the ARMS-PCR method and had a strong correlation between the marker and SR (p-value of AA = 0.012; AG = 0.359, and GG = 0.001). The resistant population has significantly higher ALF and SOD gene expression levels and AA genotype frequency. The SNP marker was inherited, so the broodstock and pedigree have the same genotype frequencies according to chi-square analysis (χ2 = 0.46 and p-value = 0.497). These results suggested that the g.455 genotype AA could be selected to produce WSSV-resistant Pacific white shrimp.

Association of IFN-γ +874 A/T SNP and hypermethylation of the -53 CpG site with tuberculosis susceptibility.

Tuberculosis (TB) is now the 2nd leading infectious killer after COVID-19 and the 13th leading cause of death worldwide. Moreover, TB is a lethal combination for HIV-patients. Th1 responses and particularly IFN-γ are crucial for immune protection against Mycobacterium tuberculosis infection. Many gene variants for IFNG that confer susceptibility to TB have been described in multiple ethnic populations. Likewise, some epigenetic modifications have been evaluated, being CpG methylation the major epigenetic mark that makes chromatin inaccessible to transcription factors, thus avoiding the initiation of IFNG transcription. We evaluated both genetic and epigenetic changes involved in IFN-γ production and TB susceptibility in Argentine population. Amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) was performed for the IFN-γ +874 A/T polymorphism (rs2430561) genotyping in 199 healthy donors (HD) and 173 tuberculosis (TB) patients. IFN-γ levels from M. tuberculosis-stimulated PBMCs were measured by ELISA. The methylation status at the -53 CpG site of the IFNG promoter in individuals with latent infection (LTBI), TB and HD was determine by pyrosequencing. Using a case-control study, we found that A allele and, consequently, AA genotype were overrepresented in patients with active disease. Moreover, HD carrying T allele (AT or TT genotype) evidenced an augmented IFN-γ secretion compared to TB patients. Codominance was the genetic model that best fits our results according to the Akaike information criterion (AIC). In addition, increased methylation levels at the -53 CpG site in the IFN-γ promoter were observed in whole blood of patients with active TB compared to LTBI individuals. IFN-γ is regulated by genetic variants and epigenetic modifications during TB. Besides, AA genotype of the rs2430561 single nucleotide polymorphism could be considered as a potential TB susceptibility genetic biomarker in Argentina and the methylation of the -53 CpG site could result in a useful predictor of TB reactivation.

Src homology 2-B adapter protein 3 C784T and Methylenetetrahydrofolate reductase C677T Polymorphisms and Inflammation Markers in ST-segment Elevation Myocardial Infarction Patients.

Neutrophil-lymphocyte (NLR), platelet-lymphocyte (PLR), eosinophil-lymphocyte (ELR), and monocyte-lymphocyte (MLR) ratios are systemic inflammatory markers related to myocardial infarction. The aim of this study is to investigate the association of Src homology 2-B adapter protein 3 (SH2B3) C784T and methylenetetrahydrofolate reductase (MTHFR) C677T polymorphisms (SNP) with systemic inflammatory markers and the severity of coronary artery disease (CAD) in 150 ST-elevation myocardial infarction (STEMI) patients. Single nucleotide polymorphisms were genotyped using the tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR) method. The inflammatory markers were calculated. An interventional cardiologist blinded to other data assessed the SYNTAX (SX) Score. Eosinophil and platelet counts were significantly higher in SH2B3 variants than in the wild type. Additionally, SH2B3 variants had significantly higher ELR than the wild type (.12 ± .19 vs .25 ± .34, p = .018). NLR, PLR, ELR, and MLR were considerably higher in MTHFR variants than in the wild type (p < .05). The SX score was significantly higher in both SH2B3 C784T (21.24 ± 8.90 vs 15.29 ± 9.40, p = .00) and MTHFR C677T (20.34 ± 10.21 vs 16.08 ± 8.39, p = .00) variants when compared with wild type. In conclusion, these polymorphisms are associated with several markers of systemic inflammation as well as the severity of CAD.

Association of ANRIL Gene Single-Nucleotide Polymorphisms With Allergic Rhinitis in Kurdish Population From Kermanshah, Iran.

Allergic rhinitis (AR) is the most common inflammatory disorder of the upper airway caused by aberrant immune responses to allergens in genetically predisposed individuals. Recently, the long noncoding RNA (lncRNA) antisense noncoding RNA in the INK4 locus (ANRIL) has been identified as a novel genetic factor associated with increased AR risk. This study aimed to evaluate the potential correlation of ANRIL gene single nucleotide polymorphisms (SNPs) with AR risk in the Kurdish population of Kermanshah, Iran. In this case-control study, 130 AR patients and 130 healthy controls were recruited to genotype for two SNPs of the ANRIL gene (rs1333048 and rs10757278) using the Tetra-primer amplification refractory mutation system polymerase chain reaction (T-ARMS-PCR) method. Our results showed no significant difference for the alleles and genotypes frequency distribution of lncRNA ANRIL SNPs (rs1333048 and rs10757278) between AR patients and healthy controls (p > 0.05). Additionally, the dominant, additive and recessive genetic models of both SNPs were not associated with altered susceptibility to AR risk (p > 0.05). The results demonstrated that the ANRIL gene rs1333048 and rs10757278 polymorphisms might not be associated with susceptibility to AR in the Kurdish population of Kermanshah, Iran.

Immunogenic Potential of the Mediterranean Fever Gene in Patients with Coronavirus Disease: A Cross-Sectional Study.

In December 2019, an outbreak of pneumonia caused by the novel coronavirus disease 2019 (COVID-19) became a pandemic and caused a global health crisis. This study evaluates the immunogenic potential of the Mediterranean fever (MEFV) gene in patients with COVID-19. A cross-sectional study was conducted from March to April 2020 in various COVID-19 referral centers in Ardabil, Iran. Blood samples of 50 hospitalized patients with confirmed COVID-19 were evaluated for MEFV gene mutation using the amplification refractory mutation system polymerase chain reaction (ARMS-PCR) and Sanger sequencing. Statistical analysis was performed using SPSS software, version 22.0. Mutations of the MEFV gene were found in 6 (12%) of the patients. All mutations were heterozygous, and no homozygous or compound heterozygous forms were detected. The total mutant allele frequency was 6% and the carrier rate was 12%. The most common allele of the MEFV variant was E148Q, detected in 3 (6%) patients. No mutant variant of the MEFV gene was detected in deceased patients. None of the mutation carriers had familial Mediterranean fever (FMF) symptoms or a family history of FMF. MEFV gene mutations may have immunogenic potential in patients with COVID-19. A preprint version of this article has already been published at https://www.researchsquare.com/article/rs-69373/latest.pdf.

Exploring the Associations and Molecular Impacts of miR-146a/rs2910164 and miR-196a2/rs185070757 with Rheumatoid Arthritis in a Pakistani Population

Introduction: MicroRNAs (miRNAs) are a new class of molecules that participate in post-transcriptional regulation of gene expression and hence have been reported to have a crucial role in the pathogenesis of rheumatoid arthritis (RA). We aimed to investigate the association of miR-146a rs2910164 (G/C) and miR-196a2 rs185070757 (T/G) with RA susceptibility in Pakistani patients and to bioinformatically predict the molecular function of these miRNAs. Methods: A case-control study on 600 individuals was conducted, including 300 RA patients and 300 matching healthy controls. Genotyping was performed by tetra-primer amplification of refractory mutation system-polymerase chain reaction, and the association between variants and RA was statistically determined using different models. Results: For the variant rs2910164 (G/C) in miR-146a, no difference in genotype distribution was observed between RA cases and controls, as determined using co-dominant (χ² = 4.33; p = 0.114), homozygous dominant (C/C vs. G/G + C/G) (OR = 0.740 [0.531–1.032]; p = 0.091), homozygous recessive (G/G vs. C/C + G/C) (odds ratio [OR] = 01.432 [0.930–2.206]; p = 0.126), heterozygous (G/C vs. C/C + G/G) (OR = 1.084 [0.786–1.494]; p = 0.682), and additive (OR 0.778 [0.617–0.981]; p = 0.039) models. Similarly, the GT genotype in the rs185070757 (T/G) miR-196a2 variant did not differ between cases and controls with any models (p > 0.05). For the first time, we report no association of miR-146a rs2910164 (G/C) and miR-196a2 rs185070757 (T/G) with RA in a Pakistani population. A subsequent bioinformatic analysis revealed that the CC genotype of miR-146a rs2910164 might have a protective role against RA pathogenesis, with no effect observed with the miR-196a2 rs185070757. Conclusion: Our findings suggest that these miRNAs might have little-to-no impact on the RA pathogenesis in the Pakistani population.

Interakcija genetskih varijanti IL-17A (rs2275913-G/A) i IL-17F (rs763780 T/C) sa aknama vulgaris

Objective. Acne vulgaris is a prevalent dermatological disorder with a complex etiology that affects pilosebaceous units and manifests both as inflammatory and noninflammatory skin conditions. The present preliminary study was aimed at evaluating the expression of IL-17A and IL-17F gene polymorphisms in acne vulgaris patients and their relationship to the severity of the condition. Methods. Sixty acne vulgaris patients and 60 healthy individuals (control group) were recruited for the study. Demographic information was obtained from the participants. Also, blood samples were collected and used to isolate DNA. The detection of IL-17A and IL-17F gene polymorphisms was assessed using tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). Results. There was no significant (p &gt; 0.05) difference between patient and control groups in the correlation between the IL-17A gene polymorphism and acne vulgaris disease in all codominance and recessive modes, but the AA and G/A genotypes can be regarded as risk factors (OR = 2.77 and 2.19, respectively) concerning the etiological fraction (EF = 0.346 and 0.331, respectively). Also, there was no statistically significant (p = 0.275) difference between the two groups in terms of the genotypes and allele frequencies for the IL-17F SNP. However, the risk analysis revealed that the CC (OR = 3.42) and T/C genotypes (OR of 1.37) may be taken into account as risk factors and etiological fractions for acne vulgaris. Conclusion. This preliminary study suggests that AA and G/A genotypes in IL-17A and CC and T/C genotypes in IL17F may be risk factors for acne vulgaris.

Correlation Analysis of Ki67 Expression and EGFR Mutation on the Risk of Recurrence and Metastasis in Postoperative Patients with Stage I Lung Adenocarcinoma

The prognosis of stage I non-small cell lung cancer (NSCLC) is generally good. However, some of the stage I NSCLC patients still may have early recurrence and metastasis, and there is no standard method to screen this part of the population. The aim of this study is to investigate the relationship between Ki67 expression as well as epidermal growth factor receptor (EGFR) mutation and the risk of recurrence in postoperative patients with stage I lung adenocarcinoma. We retrospectively enrolled 118 postoperative patients with stage I lung adenocarcinoma. EGFR mutation was tested using amplification refractory mutation system polymerase chain reaction (ARMS-PCR) , and Ki67 level was detected by immunohistochemistry (IHC), followed by the collection of the patients' clinical characteristics. Kaplan-Meier method, Log-rank test, and Cox proportional hazards regression model were used for the prognostic statistical analysis. Among the 118 patients, the rate of high Ki67 expression was 43.22% (51/118), which is related to gender, smoking status, surgical method, differentiation degree, and postoperative stage (P<0.05). Meanwhile, EGFR mutation rate was 61.02% (72/118), of which EGFR exon 19 deletion mutation rate was 19.49% (23/118), and the EGFR exon 21 L858R mutation rate was 41.53% (49/118). However, Ki67 expression was not associated with EGFR mutation status (χ2=1.412, P=0.235). Survival analysis showed that high Ki67 expression was inversely associated with disease-free survival (DFS) and overall survival (OS) in stage I lung adenocarcinoma (P<0.05), but EGFR mutation status was not significantly associated with DFS and OS (P>0.05). In the subgroup analysis, the DFS of the EGFR exon 19 deletion group was significantly decreased compared with the EGFR exon 21 L858R mutation group (P=0.031), but there was no significant difference in OS (P=0.308). Multivariate analysis showed that there was statistical significance between Ki67 expression (P=0.001) and DFS in stage I lung adenocarcinoma; Ki67 expression (P=0.03) and gender (P=0.015) were associated with OS in stage I lung adenocarcinoma. Ki67 expression is an independent influencing factor for postoperative recurrence and OS of stage I lung adenocarcinoma and it is not significantly associated with EGFR mutation. There is no significant difference between EGFR mutation status and the prognostis of stage I lung adenocarcinoma. However, the prognosis differed in EGFR mutation types; the patients with EGFR exon 19 deletion are at higher risk of recurrence than EGFR exon 21 L858R mutation.

The Evaluation Of rs11776042 Polymorphism Effect On Colorectal Cancer Risk In The Iranian Population: A Case-Control Study

Background: Recently, it has been shown that, piwi-interacting RNAs (piRNAs) as a new class of non-coding RNAs (ncRNAs), play crucial roles in germline development and carcinogenesis. Despite this, the study on the effects of piRNAs polymorphism (piR-SNP) on colorectal cancer (CRC) risk is scarce. We evaluate the impact of rs11776042 in piRNA 015551 on CRC initiation and development in the Iranian population for the first time.&#x0D; Methods: The association of novel polymorphisms rs11776042 in piRNA 015551 gene with CRC risk using a case-control study on the Iranian population was estimated. In this project 284 CRC patients and 389 non-cancerous controls were evaluated by TETRA primer-Amplification refractory mutation system polymerase chain reaction (TP-ARMS- PCR assay).&#x0D; Results: The genotypes frequency was 27%, 68% and 0.05% for C/C, C/T and T/T in controls and 31%, 65% and 0.04% in CRC patients respectively. The frequency of the C allele was 63% in patients versus 61% in controls and, T allele frequency was 37% in patients versus 39% in controls.&#x0D; Conclusion: No significant difference was found in genotype and allele frequencies between the cases and controls for rs11776042 polymorphism in piRNA 015551 in our population.

Association of the ANGPTL3 gene polymorphisms and haplotypes with cardiovascular diseases in Birjand longitudinal aging study (BLAS)

SubjectCardiovascular disease is now well established as an interaction between genetic and environmental components. Newly identified single nucleotide polymorphisms of angiopoietin-like 3 (ANGPTL3) influence lipid concentrations and risk of coronary artery disease. The current study aimed to determine the association between ANGPTL3 gene variants with incident CVDs in elderly population of the Birjand longitudinal aging study (BLAS).MethodTotally, 360 individuals were recruited in baseline of BLAS including 128 patients with CVD and 153 control subjects. DNA extraction of samples and genotyping were performed by Tetra-ARMS PCR (amplification refractory mutation system polymerase chain reaction). The association between three polymorphisms of ANGPTL3 gene (rs1748195, rs11207997, and rs10789117 variants) with CVD and its risk factors were evaluated using multivariate analysis.ResultsUnivariate and multiple analyses showed that individuals carrying the GG genotype of rs1748195 and those carrying the TT genotype of rs11207997 directly increased the risk of CVD. CC genotype of rs1748195 and rs11207997 polymorphisms had a significant negative relationship with the disease. In addition, the findings of this study indicate a significant difference in LDL, HDL, cholesterol levels between different genotypes of the rs1748195 and rs10789117 in the healthy group. Individuals with haplotypes CAC, CCC and CCT showed a significant positive relationship with CVD, CVA, AMI and CHD. As well as haplotype was associated with a 1.7-fold increase in risk of CVA, AMI, and CHD.ConclusionWe found that polymorphism of ANGPTL3 gene might support to identify individuals with a cardiometabolic and genetic disorders susceptibility. Three haplotypes CAC, CCC and CCT associated with CVD, CVA, AMI and CHD were reported.

Impact of daily vitamin D3 supplementation on the risk of vitamin D deficiency with the interaction of rs2282679 in vitamin D binding protein gene (GC) among overweight and obese children and adolescents: A one-year randomized controlled trial.

The rs2282679 polymorphism in the vitamin D binding protein (DBP) gene may influence the response to vitamin D supplementation. Therefore, we examine the effect of 1-year vitamin D supplementation on vitamin D deficiency (VDD) with the interaction of rs2282679 polymorphism in overweight and obese children and adolescents. The participants (n = 300) were part of a randomized controlled trial who received a daily supplement of either 1,000 or 2,000 IU or four supplements of 1,000 IU weekly (equal to 600 IU daily) of vitamin D3 for 12 months. Genotyping was performed using amplification refractory mutation system polymerase chain reaction (ARMS-PCR). The mean of 25(OH)D values at baseline for participants with the TT, TG, and GG genotypes were 15.4, 14.4, and 10.8 ng/mL, respectively, and were not different between the three genotype groups (P = 0.062). A significant reduction in VDD was observed after vitamin D supplementation with dosages of 1,000 or 2,000 IU compared to 600 IU. No significant association of genotypes with risk of VDD was observed in each intervention group after vitamin D supplementation, except, that individuals with TG genotype showed a higher risk of VDD compared to those with TT genotype in the 2,000 IU group after 6 months of supplementation [odds ratio (95% CI): 6.94; 1.30-37.02]. We observed no interaction between time duration, three genotypes, and dosages with serum 25(OH)D, calcium, phosphorus, alkaline phosphatase, and parathyroid hormone levels. Response to vitamin D supplementation by three doses of 600, 1,000, and 2,000 IU could not be affected by rs2282679 polymorphism during 12 months in overweight and obese children and adolescents.