majid motovali-bashi

Various mutations in factor VIII locus are led to an X-linked bleeding disorder in hemophilia A. One of the leading causes of inefficient remedy for hemophilia A disease is the lack of specific and sensitive procedure for punctual diagnosis of the disease. miRNAs indicate that they have a great potential as diagnostic and prognostic biomarkers due to their stability in body fluids and their capability to identification. Furthermore, a relationship between alteration in expression levels of miRNAs and onset and progression of the hemophilia A disease has been reported.Prediction of new miRNAs and nomination as regulators of FVIII gene has been considered as the objective of this study. The ability to express new miRNAs in FVIII locus was studied via reliable bioinformatic databases such as SSCprofiler, RNAfold, miREval, FOMmiR, MaturBayes, miRFIND, UCSC genome browser, Deep Sequencing, and miRBase. Output data of previously mentioned databases were analyzed, and one stem-loop structure was qualified, and the region is predicted to express new miRNA. The presented stem-loop structure can be used as biomarker in diagnosis of the disease and regulation of factor VIII gene after subsequent experimental verification.

Some previous human and animal studies have supported the idea that KDM3A down-regulation might be the main cause of male infertility, especially in non-obstructive azoospermia (NOA). The regulatory role of micro-RNAs (miRNA) has been investigated in the development of male infertility.

The expression level of hsa-miR-30a-5p in azoospermia was evaluated to reveal its possible association with the etiology of male infertility.

In this case-control study, 30 men with azoospermia (19 of whom had NOA) were selected as the case individuals, and 11 men with obstructive azoospermia (OA) were selected as control individuals. The best miRNA with the strongest ability to target the KDM3A gene was detected via comprehensive bioinformatics analysis. Reverse transcriptase quantitative polymerase chain reaction was used to assess the expression level of hsa-miR-30a-5p. After analyzing the data, the expression level of hsa-miR-30a-5p was compared between men with NOA and men with OA.

The findings supported the idea that hsa-miR-30a-5p is the miRNA with the best ability to target the KDM3A transcript. The expression analysis of hsa-miR-30a-5p indicated a significant overexpression (p = 0.04) in men with NOA compared to in men with OA.

Hsa-miR-30a-5p was overexpressed in men with NOA compared to in control individuals. Hsa-miR-30a-5p could target the KDM3A transcript and may suppress its expression.

Hemophilia-A is a common genetic abnormality resulted from decreased or lack of factor VIII (FVIII) pro-coagulant protein function caused by mutations in the F8 gene. Majority of molecular studies consider screening of mutations and their relevant impacts on the quality and expression levels of FVIII. Interestingly, some of the functions in FVIII suggest a probable involvement of small non-coding RNAs embedded within the sequence of F8 gene. Therefore, microRNAs which are encoded within the F8 gene might have a role in hemophilia development. In this study, miRNAs production in the F8 gene was investigated by bioinformatics prediction and experimental validation.

In this experimental study, bioinformatics tools have been utilized to seek the novel microRNAs inserted within human F8 gene. The ability to express new microRNAs in F8 locus was studied through reliable bioinformatics databases such as SSCProfiler, RNA fold, miREval, miR-FIND, UCSC genome browser and miRBase. Then, expression and processing of the predicted microRNAs were examined based on bioinformatics methods, in the HEK293 cell lines.

We are unable to confirm existence of the considered mature microRNAs in the transfected cells.

We hope that through changing experimental conditions, designing new primers or altering cell lines as well as the expression of vectors, exogenous and endogenous expressions of the predicted miRNA will be confirmed.

Hemophilia A is an X-linked bleeding disorder resulting in a deficiency of plasma clotting factor VIII and caused by mutations in the FVIII gene (F8 gene). MicroRNAs (miRNAs) in body fluids are promising biomarker candidates for Hemophilia A, due to their stability in body fluids and accessibility by non- or minimally-invasive procedures. Therefore; Advances in miRNA analysis methods resulted in a wide range of publications on miRNAs as putative biomarkers.

Here we tried to scan the F8 gene region to predict a novel miRNA and identify it as a regulator of the F8 gene.

To this aim, the ability to express novel miRNAs in F8 locus was assessed via reliable bioinformatics databases such as SSCprofiler, RNAfold, miREval, FOMmiR, MaturBayes, miRFIND, UCSC genome browser, Deep Sequencing, and miRBase.

Data analysis from the relevant databases offers one stem-loop structure that is predicted to express a novel miRNA.

The diagnosis of Hemophilia A with the help of these types of biomarkers is a non-invasive procedure that has been demonstrated to have a significant role in the early diagnosis of the disease. Hopefully, the proposed candidate sequence will be confirmed in vitro and become a non-invasive biomarker in the near future.

Hemophilia A is an X-linked bleeding disorder that occurs due to the deficiency of Factor VIII (FVIII) protein clotting activity. The mutations in the F8 gene, which encodes FVIII coagulating protein have been widely reviewed. However, there is a wide range of criteria that in addition to F8 gene mutations, different molecular mechanisms may be associated with hemophilia A. Various functions of FVIII could be related to the hypothetical small non-coding RNAs, located within the F8 gene sequence. Therefore, miRNAs that can post-transcriptionally regulate gene expression might confer susceptibility to developing hemophilia A. Here, we have selected a bioinformatically predicted hairpin structure sequence in the first intron of the F8 gene that has the potential to produce a real miRNA (named put-miR1). We tried to experimentally detect the predicted miRNA via RT-PCR following its precursor overexpression in HEK 293 cell lines. Despite the accuracy of miRNA prediction, according to the reliable bioinformatics studies, we couldn’t confirm the existence of considered mature miRNA in transfected cells. We hope that through changing experimental conditions, designing new primers, or altering cell lines and expression vectors, the exogenous and endogenous expression of the predicted miRNA will be confirmed.

The role of KDM3A and its downstream genes in male fertility has been approved in animal models. Additionally, the expression shrinkage of KDM3A is significantly correlated with human azoospermia phenotype. Aberrant expression of micro-RNAs could mislead spermatogenesis and mostly lead to diverse phenotypes of male infertility.

The aim of this study was to evaluate the expression level of hsa-miR-27a-3p in azoospermic men to reveal its possible association with infertility.

This case-control study was conducted on 30 azoospermic men, of whom, 19 had non obstructive azoospermia (NOA) and 11 obstructive azoospermia (OA) according to the pathological examinations. Comprehensive bioinformatics investigations were performed securely and hsa-miR-27a-3p was selected afterward. Reverse Transcriptase-quantitative polymerase chain reaction (RT-qPCR) method was used and statistical analysis was performed to compare the expression level of hsa-miR-27a-3p in both OA and NOA individuals.

In silico analysis suggested hsa-miR-27a-3p, with its potential binding ability to target KDM3A transcripts. The expression analysis of candidate hsa-miR-27a-3p indicated its significant overexpression in NOA men.

The hsa-miR-27a-3p was overexpressed in NOA men compared to OA-control individuals. As a consequence, the overexpressed micro-RNA could downregulate directly KDM3A and indirectly TNP1 and PRM1. Therefore, spermatogenesis could be misled and male infertility could be developed.

Various mutations in factor VIII (F8) gene locus are led to an X-linked bleeding disorder in patients with hemophilia A. One of the leading causes of inefficient treatment available for hemophilia A is the lack of specific and sensitive diagnostic procedure for the disease. The discovery of a functional role of microRNAs (miRNAs) in the pathogenesis of a wide range of human diseases makes them the potential, non-invasive, biomarker candidates for hemophilia A. Therefore, advances in computational tools for miRNA discovery leads to numerous recent publications on miRNAs as putative biomarkers.

The current study aimed at scanning the F8 gene region to predict novel miRNAs as regulators of the F8 gene.

The potential of the FVIII locus to express new miRNAs was studied via reliable bioinformatics databases, such as SSCprofiler, RNAfold, miREval, miR-Find, FOMmiR, UCSC genome browser, and miRBase.

Data analysis from previously mentioned databases offered two stem-loop structures predicted to express novel miRNAs.

The presented stem-loop structures can be used as powerful non-invasive biomarkers in early diagnosis of the disease and regulation of the factor VIII gene after subsequent experimental verification.

A major complication in treating hemophilia A is the development of neutralizing antibodies (inhibitors) against therapeutic administered factor VIII (FVIII), which occurs in approximately 20-30% of patients with severe disease. These inhibitors render FVIII replacement therapy ineffective and increase the morbidity and mortality risk. The currently accepted method to eradicate inhibitors is immune tolerance induction (ITI), and frequent intensive administration of FVIII until inhibitor titers drop. Current ITI protocols are extremely costly and not effective in all patients. During the last decade, many types of research have been accomplished to clarify the mechanisms that mediate immune tolerance induction. Novel experimental therapies including monoclonal antibodies, viral vector-mediated gene therapy, regulatory T cell induction using immunosuppressive drugs, and nanoparticle-based immune modulation show promising results in hemophilia A clinical trials. This review focuses on treatment options towards the anti-FVIII immune responses and current novel therapies in clinical trials.

Tumor-infiltrating regulatory T (TI-Treg) cells perform the significant function in cancer immune escape. In this study, the third generation CAR construct was designed against human CD25 antigen, the significant cell surface biomarker of TI-Tregs.

Initially, the construct of anti-CD25 CAR was designed. Using RNAfold web server, the RNA secondary structure was evaluated. Also, utilizing lentiviral vectors, NK-92 cell line was transduced. Afterward, the expression level of anti-CD25 CAR RNA was assessed by qRT-PCR in NK-92 cells transduced with CAR and mock transfer vectors and also untreated cells.

The RNA secondary structure was stable. Also, the expression level of anti-CD25 CAR RNA in transduced NK-92 cells by pCDH-513B-1-anti-CD25 CAR transfer vector was significantly higher than transduced NK-92 cells by mock transfer vector and untreated cells (p˂0.0001).

The present study on anti-CD25 CAR RNA showed that this type of CAR transcripts were stable and expressed at high level. In fact, this type of CAR can be further studied in the future as a tool to remove the cancer immune escape in all types of solid and liquid cancers.

Breast cancer is one of the major causes of illness and mortality among women. Long non-coding RNAs (LncRNAs) have an important role in tumor development and progression. MALAT1 (Metastasis-associated lung adenocarcinoma transcript 1) is a lncRNA that deregulates in several cancers, however, its value in the diagnosis of breast cancer is unclear. This study was conducted to investigate the MALAT1 expression levels in breast tumor tissue, its association with tumor clinical features and its diagnostic value as a biomarker in breast cancer. In this study, the expression level of MALAT1 was measured in 31 breast tumor tissues and 31 adjacent normal tissues by real-time polymerase chain reaction. The MALAT1 expression alteration between the tumor and normal tissues and its association with clinical characteristics were analyzed with t-test and one-way ANOVA, respectively. The MALAT1 role as a biomarker was investigated by ROC curve. MALAT1 expression level in tumor tissues than to adjacent normal tissues showed a significant decrement 2.87 times (fold change= 0.348, p<0.001). Also, there was a positive and significant correlation between MALAT1 expression and patient's age and tumor invasive features (p <0.05). The ROC curve results showed that the area under the curve was significant and equal to 0.773. The sensitivity and specificity of MALAT1 as biomarker were 71.43 and 91.67, respectively. The results of this study showed that the MALAT1 has a significant reduction in breast cancer and can act as a biomarker in diagnosing patients from healthy individuals.

The eukaryotic replisome is a multi-component complex that drives DNA replication with a speed of approximately 2 to 3 kb per min. This implies that chromatin is disrupted at a rate of around 10 to 15 nucleosomes every minute a head of each active replisome. To reproduce a similar chromatin environment on new DNA, histones and other chromatin-bound factors are transferred from the parental strand to the daughter strands. In addition, new histones are incorporated to maintain nucleosome density, and their modification signature should be assimilated to nearby old histones in the local chromatin environment. In general interaction between the components of the replisome and chromatin proteins can help to understand the proper way of improving the replication fork and its relationship to chromatin. In this study, the regulatory mechanism of chromatin replication and epigenome maintenance are evaluated.

Beta-thalassemia is one of the most prevalent inherited blood diseases among Iranians. The aim of this study was to elucidate the chromosomal background of beta-thalassemia mutations in Esfahan province, Iran. In this study, we investigated three frequent mutations (c.315+1G>A, c.93-21G>A and c.92+5G>C in the β-globin gene, the frequency of RFLP haplotypes, and LD between markers at β-globin gene cluster) in 150 beta-thalassemia patients and 50 healthy individuals. The molecular and population genetic investigations were performed on RFLP markers HindIII in the c.315+1G>A of Gγ (HindIIIG) and Aγ (HindIIIA) genes, AvaII in the c.315+1G>A of β-globin gene and BamHI 3' to the β-globin gene. All statistical analyses were performed using Power Marker software and SISA server. Fifty percent of beta-thalassemia patients were associated with these mutations. Haplotype I was the most prevalent haplotype among beta-thalassemia patients (39.33%) and normal individuals (46%). The commonest c.315+1G>A mutation in our population was tightly linked with haplotype III (43.75%) and haplotype I (31.25%). The second prevalent mutation, c.92+5G>C, was 90%, 6.66%, and 3.33% in linkage disequilibrium with haplotypes I, VII, and III, respectively. The c.93-21G>A mutation indicated a strong association with haplotype I (80%). Our study participants like beta-thalassemia patients from Kermanshah province was found to possess similar haplotype background for common mutations.The emergence of most prevalent mutations on chromosomes with different haplotypes can be explained by gene conversion and recombination. High linkage of a mutation with specific haplotype is consistent with the hypothesis that chromosomes carrying beta-thalassemia mutations experienced positive selection pressure, probably because of the protection against malaria experienced by beta-thalassemia carriers.

Colorectal cancer is the third cancer which results in death in western countries. Some of the risk factors of this cancer are age, inadequate diet, obesity, inactivity, genetic changes, etc. Considering the relationship between genetic changes with colorectal cancer is the subject of recent studies, and one of the subjects which have been focused is T241M polymorphism in Xrcc3 gene.
In this case-control study, 90 patients with colorectal cancer and 83 healthy people were included. Genomic DNA was extracted from peripheral blood, and genotype distribution in the region of polymorphism was studied. By designing of primers, the considered area was amplified and genotyped using polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) method.
Findings: We observed the association between T241M polymorphism and colorectal cancer. Besides, there was a meaningful relationship between aging and family history with this cancer. On the other hand, smoking was not associated with colon cancer, but it also showed a significant role in the incidence and metastasis of rectal cancer.
The relationship between T241M polymorphism and increased risk of colorectal cancer, along with family history of cancer, old age, and smoking were reported as risk factors for colorectal cancer. Therefore, the results of this research can show the need for further investigation in subsequent studies.

One third of male factors related to infertilities are basically unknown and it has been predicted to be most of the unknown infertilities based on genetic abnormalities. On the other hands, the effect of HLA-DRA genes on spermatogenesis has been shown recently. So, this study was aimed to assess the association of rs7192 polymorphic site of HLA-DRA gene on male infertility in Isfahan province of Iran.
100 infertile men with azoospermia or severe oligozoospermia as case group and equal numbers of fertile men as control individuals were selected. Genomic DNA of peripheral blood was extracted after the written consent was taken. rs7192 polymorphic site has been analyzed by ARMS-PCR method in the case and control groups.
According to the results, the frequency of G allele was higher in infertile men than the T allele; but, allele frequency differences was not significant between the case and control groups (OR=0.86, P=0.47). However, it has been shown that GG homozygous in comparison to TT homozygous show an increased risk of male infertility but it was not statistically significant (OR=1.47, P=0.4). In parallel, comparison of GG GT genotypes to TT ones was not also statistically significant between infertile and fertile men (OR=1.43, P=0.4).
Association study of polymorphic rs7192 did not show any significant relationship among men infertile with azoospermia or severe oligozoospermia in Isfahan province of Iran.

Allergic rhinitis, a chronic inflammatory disease of the upper airways, has a major effect on the life quality of patients. The inflammatory response in the nasal mucosa includes an immediate IgE-mediated response by mast cell, so that the late phase of response is characterized by recruitment of eosinophilsand basophils. IL-18 is a member of the IL-1 family and originally is described as an IFN-γ –inducing factor (IGIF), also it is known for influencing the balance of Th1/Th2 immune response. The aim of this study to investigated that whether immunoglobulin (Ig) E levels in serum are associated with allergic rhinitis.
Genotyping for -137G/C SNPs of IL18 promoter was performed using 130 patients with AR and 62 healthy control volunteers then serum levels of total IgE were determined by ELISA method. Statistical analyses were carried out by use of SPSS version 19.
The results showed that no significant relationship between genotypes -137G / C polymorphism and serum IgE levels between patients and controls in allergic rhinitis (P> 0.05).
According to the results of this study, it seems that the IL18 gene polymorphism -137G/C is not associated with IgE levels and susceptibility of allergic rhinitis

Breast cancer is the most common cancer among women. Polymorphisms of micro RNA genes such as microRNA 152 and microRNA 148a that they are involved in cell proliferation, angiogenesis and apoptosis could be a potential factor for increasing risk of breast cancer and its development. So, the aim of this study is to investigate the association between polymorphisms of these micro-RNAs and age of onset and progression of breast cancer.
This case-control study was conducted on 100 patients of breast cancer and 100 healthy women. The routine clinical tests were performed in studied patients regularly. After genotyping using RFLP-PCR and Tetra primer ARMS-PCR techniques, subjects of this study were categorized into three genotypes including AA,AG and GG for the miR-148a polymorphism and TT,TC and CC for the miR152 polymorphism. The collected data through the SISA website and chi-square tests were analyzed.
Findings: The results show that the miR-148a and miR152 polymorphisms reduce age of onset of cancer, but this is not statistically significant (miR152: OR=2.16, P=0.06), (miR-148a: OR=2.54, P=0.06). There is also no significant association between these polymorphisms and stages of cancer progression were observed (miR148a: OR=1.16, P=0.09); )miR152: OR=0.47, P=0.13).
It seems that the G and T alleles can be as a risk factor for the reduction of age of onset and breast cancer progression in Isfahan population. But to determine the alleles as biology risk factors further studies and more research is needed.

Infertility is a health problem which affects about 10-20% of married couples. Male factor infertility is involved approximately 50% of infertile couples. Most of male infertility is regarding to deletions in the male-specific region of the Y chromosome.

In this study, the occurrence of deletions in the AZF region and association between infertility and paternal age were investigated in Iranian men population.

To assess the occurrence of Y chromosomal microdeletions and partial deletions of the AZF region, 100 infertile men and 100 controls with normal spermatogenesis were analyzed. AZFa, AZFb, AZFc and partial deletions within the AZFc region were analyzed using multiplex PCR method. Finally, the association between paternal age and male infertility was evaluated.

No AZFa, AZFb or AZFc deletions were found in the control group. Seven infertile men had deletions as the following: one AZFb, five AZFc, and one AZFab. Partial deletions of AZFc (gr/gr) in 9 of the 100 infertile men (9/100, 9%) and 1 partial AZFc deletions (gr/gr) in the control group (1/100, 1%) were observed. In addition, five b2/b3 deletions in five azoospermic subjects (5/100, 5%) and 2 partial AZFc deletions (b2/b3) in the control group (2/100, 2%) were identified. Moreover, the risk of male infertility was influenced by the paternal age.

The results of this study suggested that the frequency of Y chromosome AZF microdeletions increased in subjects with severe spermatogenic failure and gr/gr deletion associated with spermatogenic failure.

β-thalassemia is the most common monogenic disorder in human. The (CT) polymorphism at -158 upstream region of the γG-globin gene and pharmacological factors such as hydroxyurea have been reported to influence γ-globin gene expression and the severity of clinical symptoms of β-thalassemia. In the present study, 51 β-thalassemia intermediate patients were studied. Xmn1γG polymorphism genotype was determined using Tetra-Primer ARMS-PCR technique. Hemoglobin (Hb) and fetal hemoglobin (HbF) levels were determined by gel electrophoresis. Of 51 patients, 35 (68.6%) patients were heterozygous (CT) and 16 (31.4%) patients were homozygous (CC). Of 30 patients under treatment by hydroxyurea, 20 (66.7%) patients were heterozygous (CT) and 10 (33.3%) patients were homozygous (CC). Our results demonstrated that in the heterozygous (CT) genotype, the Hb (9.58 ± 1.25 gm/dl) and HbF (89.30 ± 21.87) levels were significantly higher in comparison with homozygous (CC) genotype (7.94 ± 1.34 gm/dl and 70.32 ± 40.56, respectively). Furthermore, we observed that after drug usage, the Hb and HbF levels in patients with heterozygous (CT) genotype (0.7 ± 1.26 gm/dl and 5.95±14.8, respectively) raised more in comparison with homozygous (CC) genotype (0.26 ± 1.43 gm/dl and 0.8±1.31, respectively). Hb and HbF levels in the patients carrying T allele are increased significantly, and they also response to hydroxyurea treatment.

The Interleukin-18 (IL-18) is a member of the IL-1 family. It was originally described as IFN-γ –inducing factor (IGIF), and is known to influence the balance of Th1/Th2 immune response and could induce the allergic rhinitis (AR) expression. The aim of this study was to investigate whether the IL-18 promoter polymorphism (-133 G/C) were associated with AR in Chaharmahal va Bakhtiari province. In this descriptive analytical study, genomic DNA was obtained from the blood samples of 293 AR patients and 218 healthy control volunteers extracted by standard phenol chloroform method. The IL-18 polymorphism was analyzed by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis. The frequency of the GC genotype of the IL-18 gene polymorphism (-133) was greater in allergic rhinitis patients than in control (P<0.05). The rate of increaser distribution was calculated 3.69 by comparing the frequency of the G allele genotype to the genotype a carrier no allele G (CC) in both groups. This study suggests that IL-18 promoter polymorphism (-133 G/C) may be participated as a risk factor in the pathogenesis of AR.

People are usually susceptible to carcinogenic aromatic amines, present in cigarrette smoke and polluted environment, which can cause DNA damage. Therefore, maintenance of genomic DNA integrity is a direct result of proper function of DNA repair enzymes. Polymorphic diversity could affect the function of repair enzymes andthus augment the risk of different cancers. Xeroderma pigmentosum group D (XPD) gene encodes one of the most prominent repair enzymes and the polymorphisms of this gene are thought to be of importance in lung cancer risk. This gene encodes the helicase, which is a component of transcription factor IIH and an important part of the nucleotide excision repair system. Studies reveal that individuals with Lys751Gln polymorphism of XPD gene have a low repairing capacity to delete the damages ofultraviolet light among other XPD polymorphisms. In this case-control study, first Lys751Gln polymorphism was genotyped, then its association with lung cancer risk was analyzed. Genomic DNA wasextracted from the whole blood sample of 640 individuals from Iran (352 healthy individuals and 288 patients). Allele frequencies and heterozygosity of Lys751Gln polymorphism were determined using polymerase chain reaction-restriction fragment length polymorphism method. According to statistical analyses, lung cancer risk in individuals with Lys751Gln polymorphism (Odd Ratio=1.8, 95% Confidence Interval 0.848-3.819) is approximately twice as high as that of Lys/Lys genotype, however 751Gln/Gln genotype did not relate to lung cancer risk (Odd Ratio=0.7, 95% Confidence Interval 0/307-1/595). This study suggests that heterozygous polymorphism (Lys/Gln) increases the sensitivity of lung cancer risk, while homozygous polymorphism (Lys/Lys) probably decreases its risk and C allele frequency shows no remarkable increase in the patients.

Allergy is regarded as a multifactorial condition that its onset and severity are influenced by both genetic and environmental factors. Hence, identification of genetic factors involved in allergic rhinitis development and its related phenotypes is a major task in understanding the genetic background of allergic rhinitis. This study was designed to examine the association between IL-18 -607 A/C promoter polymorphism on chromosome 11q22 and allergic rhinitis. In this analytic study, genomic DNA was obtained from the blood samples of 293 patients with allergic rhinitis and 218 healthy controls by standard phenol chloroform method. The IL-18/-607 A/C polymorphism was analyzed by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis. To analyze the association between genotypes and alleles and the disease in the case group compared with the control group, X2 test was used. The frequency of the AC genotype of the IL-18/-607 A/C gene polymorphism was significantly greater in allergic rhinitis patients than in controls (p<0.05). By comparing the frequency of AA genotype with other genotypes, OR was calculated as 2.03. The results suggest that IL-18/-607 A/C polymorphism gene may be one of the factors participating in the pathogenesis of AR or its intermediary phenotypes.

Two types of Cyclooxygenase enzymes، which produce prostaglandins، are present in humans. Prostaglandins play a role in the functions of various organs including the immune system، blood circulation، and cell division. Reports indicate that the level of these enzymes is increased in patients with metastases in several malignancies، including larynx، lung، stomach، colon، and prostate. The aim of this study was to investigate the association between -765 G/C promoter polymorphism of cyclooxygenase 2 and initiation and progression of lung cancer. This is a case-control، retrospective study. Genotyping of cyclooxygenases was carried out by testing blood samples from 120 lung cancer patients and 110 controls using restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) technique. The observed numbers of each cyclooxygenase genotype were compared with that expected for a population in Hardy-Weinberg equilibrium by using a χ2 test. The significance of the differences of observed alleles and genotypes between groups was tested using the odds ratio analysis. The genotype GG of polymorphism -765G/C seen in 62. 5% of patients did not indicate a significant statistic increase in comparison with the control group (55. 45%; P< 0. 6); however، when the patients were divided into two groups i. e. those with metastases and without metastases، a weak link was identified between genotype GG of polymorphism G/C، with metastasizing cancer، (P< 0. 04). Dividing patients into sub-groups such as sex and smoking habit showed no difference. It seems that there is no association between -765 G/C polymorphism and developing of lung cancer statistically. But genotype GG of polymorphism -765G/C is linked weakly with lung cancer metastases.

The clavulanic acid regulatory gene (claR) is in the clavulanic acid biosynthetic gene cluster that encodes ClaR. This protein is a putative regulator of the late steps of clavulanic acid biosynthesis. The aim of this research is the molecular cloning of claR, isolated from the Iranian strain of Streptomyces clavuligerus (S. clavuligerus). In this experimental study, two different strains of S. clavuligerus were used (PTCC 1705 and DSM 738), of which there is no claR sequence record for strain PTCC 1705 in all three main gene banks. The specific designed primers were subjected to a few base modifications for introduction of the recognition sites of BamHI and ClaI. The claR gene was amplified by polymerase chain reaction (PCR) using DNA isolated from S. clavuligerus PTCC 1705. Nested-PCR, restriction fragment length polymorphism (PCR-RFLP), and sequencing were used for molecular analysis of the claR gene. The confirmed claR was subjected to double digestion with BamHI and ClaI. The cut claR was ligated into a pBluescript (pBs) vector and transformed into E. coli. The entire sequence of the isolated claR (Iranian strain) was identified. The presence of the recombinant vector in the transformed colonies was confirmed by the colony-PCR procedure. The correct structure of the recombinant vector, isolated from the transformed E. coli, was confirmed using gel electrophoresis, PCR, and double digestion with restriction enzymes. The constructed recombinant cassette, named pZSclaR, can be regarded as an appropriate tool for site directed mutagenesis and sub-cloning. At this time, claR has been cloned accompanied with its precisely selected promoter so it could be used in expression vectors. Hence the ClaR is known as a putative regulatory protein. The overproduced protein could also be used for other related investigations, such as a mobility shift assay.

Infertility is one of the important human problems. One of the main genetic factors of infertility is the deletions in the chromosome Y‚ which is reported in 10-15% of men with severe azoospermia and oligospermia. Three regions in azoospermia factor (AZFa), (AZFb) and (AZFc) are specified as the spermatogenetic regions. In this study we investigated the effect of changes in Yq11.223 (DAZ) region in chromosome Y on infertility. In this study the blood samples of 100 infertile men who referred to the Infertility Center of Isfahan and 100 healthy people, as the control group were taken. The genomic DNA was extracted from blood samples. The analyses were performed by applying the polymerase chain reaction (PCR) technique in AZF region of Yq11.223. In this study 70 azoospermia and 30 oligospermia patients were investigated. The deletions in AZF region of Yq11.223 were recognized in 7 azoospermia patients but there were not detected in oligospermia patients and the control group. It was also observed that the intensity of bands were changed comparing between azoospermia patients. Deletion frequency in the region of Yq11.223 was observed in 7 of 100 (7%) infertile Oligospermia and Azoospermia men. Our results confirm the effect of DAZ in the man’s infertility.

Colorectal cancer is the third cause of cancer death in western countries. Age, inadequate diet, obesity, inactivity and genetic changes are some of the risk factors of colorectal cancer. Corelation of genetic diversity in homologous recombination repair system with cancer was evaluated in many recent studies. This study was done to investigate the correlation of T241M polymorphism in Xrcc3 gene and colorectal cancers. In this cross-sectional study after collecting blood samples and extraction genomic DNA, genotype distribution of the polymorphism was determined by RFLP-PCR (Restriction fragment lengh-polymerase-Polymerase chain reaction) method. A significant corelation between T241M polymorphism with colorectal cancer was seen. Age and family history were also corelated with this cancer. Although, there was no statistically relationship between smoking status and colon cancer, but it showed correlation with rectum cancer and it has been also observed that the most occurance of metastatic activity is in the rectum. According to our study T241M polymorphism in Xrcc3 gene from homologous recombination repair systm could be a suitable factor for early diagnosis of colorectal cancer especially rectum and its co-operation with smoking status.