mehran gholamin

Esophageal carcinoma (ESCA) is one of the most common types of cancer. ESCA accounted for the sixth leading cause of cancer-related deaths globally. Most patients are diagnosed at late stages of ESCA, with distance metastasis or chemoresistance, which leads to a poor prognosis. Previous studies demonstrated lncRNA presentation and roles in ESCA cells and patients' tissue. It has been proposed that lncRNAs can be considered a new prognostic and diagnostic biomarker in ESCA. In this study, we comprehensively explored the interaction of lncRNAs with miRNAs and mRNAs of the TCGA database and proposed a novel promising biomarker with good diagnostic and prognostic values.

The public data of RNA-seq, miR-seq, and related clinical data were downloaded from the TCGA database. Differential expression analysis was conducted by “limma” in R. GO, and KEGG signaling pathways were used for enrichments. STRING database was used for PPI analysis. CE-network was constructed by the STAR database in R. Kaplan-Meier survival analysis (log-rank test), and ROC curve analysis was used to indicate the diagnostic and prognostic values of the biomarkers.

Differentially expressed data illustrated that 45.8% of the total mRNAs in the data related to ESCA patients showed increased expression and 54.2% decreased expression. The GO and KEGG pathway analysis showed that the differentially expressed mRNAs were enriched in critical biological processes. Important protein-protein interaction hubs were identified. The ceRNA network data demonstrated critical lncRNAs essential in ESCA development, including TMEM16B-AS1, AC093010.3, SNHG3, and PVT1. The data revealed that the lncRNA WDFY3-AS2, AC108449.2, DLEU2, AC007128.1, and AP003356.1 are potential diagnostic and prognostic biomarkers in ESCA patients.

Altogether, this study demonstrates lncRNA, miRNA, and mRNA interaction and mentions regulatory networks which can be considered as a therapeutic option in ESCA. In addition, we proposed potential diagnostic and prognostic biomarkers for ESCA patients.

Superparamagnetic iron oxide nanoparticles (SPIONs) have been considered promising non-invasive imaging tools in medicine. However, their high surface energy leads to NPs aggregation, while non-targeted SPIONs can cause cytotoxic effects on normal cells. In this work, we evaluated the in vitro potential of polyethyleneimine (PEI)-SPIONs targeted by PNC27 peptide as a double targeting agent throughout early cancer diagnosis.

Initially, PEI was conjugated to PNC27 with HDM-2-binding domain. Then, SPIONs were loaded into PEI-PNC27 through the ligand exchange method. The physicochemical characteristics of the synthesized NPs were evaluated. The cytotoxicity and targeting efficiency were assayed against HT-29 and CT-26 cell lines along with NIH-3t3 as normal cells by MTT method and Prussian blue staining test, respectively. 

The mean diameter of synthesized carriers was obtained in the range of 86.6 – 116.1 nm with a positive charge. According to the cytotoxicity results, the binding and uptake abilities of the PNC27 peptide by cancer cells were significantly higher than that of the NIH-3t3 cells. However, the results were indicative of the more toxic impacts of targeted synthesized NPs against CT-26 cancer cell line when being compared with HT-29 cells, which may be caused by the different cytotoxicity mechanisms of NPs. In addition, the targeted carriers and SPIONs were present inside and around the cells with HDM-2 expression along with only a few non-targeted vectors, while displaying no appearance throughout the normal cell.

The results indicated the efficiency of targeted PEI-coated SPIONs for cancer diagnostic applications.

Gastric cancer (GC) is a malignancy cause associated with a high death rate in the world. Cancer stem cells (CSCs) are a rare immortal subpopulation of cells within tumors with characteristics of the ability to self-renew, initiate tumor, and differentiate into defined progenies as well as and high resistance to conventional therapies.  Despite the use of surgery and chemotherapy for GC therapy, there are no efficient therapeutic protocols for it to date. Therefore, rapid isolation of CSCs in order to therapeutic targets, especially immunotherapy is very important. Cancerous cell suspension isolated from patients with GC was cultured in the serum-free medium containing EGF, bFGF, LIF, and heparin under non-adherent culture conditions to generate spheres. Expression of mRNA level stemness transcription factors (OCT4, SOX2, SALL4, and Cripto-1), CD44 variable isoforms (CD44s, CD44v3, CD44v6, CD44V8-10) of spheroid-forming single cells compared with gastric normal tissue cells using real time PCR and molecules of CD44, CD54, and EpCAM as gastric CSC markers, and stemness factor Oct4 using flow cytometry, as well as tumorgenicity using subcutaneous injection of sphere-forming cells to nude mice were investigated. Few cancerous cells isolated from patients with GC were able to generate three-dimensional spheroid colonies in the serum-free medium containing EGF, bFGF, LIF, and heparin under non-adherent culture conditions, and form xenograft tumors in immunodeficient nude mice after subcutaneous injection. Spheroid-forming single cells upregulated stemness transcription factors OCT4, SOX2, SALL4, and Cripto-1 that are associated with pluripotency and self-renewal and CD44 isoforms (CD44s, CD44v3, CD44v6, CD44V8-10) compared with gastric normal tissue cells. Finally, molecules of CD44, CD54, and EpCAM as gastric CSC markers and stemness factor Oct4 were expressed in sphere-forming cells. We suggested that the sphere formation and tumorigenicity assays are two procedures, leading to the rapid isolation of cancer cells with certain stem-like properties in order to target CSCs using autologous dendritic cell therapy, especially in patients with advanced disease.

Besides the uncertainty about colorectal cancer stem cell (CCSC) markers, isolating, purifying, and enriching CCSCs to produce CCSC vaccines is highly challenging. However, allogeneic vaccines developed from CRC cell lines can provide universal, comprehensive, inexpensive, simple, and fast approach to cancer treatment. CCSCs were isolated from human CRC tissue using the in vitro sphere formation assay and then characterized through gene expression analysis, in vivo and in vitro tumor formation assay, karyotyping, and surface marker detection. Subsequently, CCSCs and two CRC cell lines (HT-29 and SW-480) were inactivated with cisplatin (CDDP) and administrated as vaccines to the three groups of athymic C57BL/6 nude mice. Afterward, tumorigenesis was challenged with HT-29 cells. The antitumor effect of vaccines was evaluated by tumor and spleen examination and immune response analysis. The cytotoxic activity of splenocytes and serum levels of TGF-β and IFN-γ were measured by Calcein-AM cytotoxicity assay and enzyme-linked immunosorbent assay (ELISA), respectively. The results of gene expression analysis showed that CCSCs are CD44+CD133-LGR5-. All vaccinations resulted in decreased tumor growth, spleen enlargement, enhanced serum level of IFN-γ and TGF-β, and increased cytotoxic activity of natural killer (NK) cells. The antitumor efficacy of the CCSC vaccine was not more than CRC cell line-based vaccines. Interestingly, the allogeneic SW-480 vaccine could effectively inhibit tumorigenesis. Despite the great challenge in developing CCSC vaccines, allogeneic vaccines based on CRC cell lines can efficiently induce antitumor immunity in CRC.

Glioblastoma primary brain tumor.Immunotherapy is a promising therapeutic adjuvant in fighting against this cancer.  Cancer/testis antigens (CTAs) are a group of tumor-associated antigens that are typically restricted to adult testis, but they are aberrantly expressed in several types of cancers, especially in advanced cancers with stem-like characteristics. These tumor-associated antigens are immunogenic in different cancers. Finding of frequently expressed CTAs in GBM can provide effective immunotherapeutic targets to use in translational researches on this cancer. The aim of this study was conduct an extensive expression analysis of ADAM29, HORMAD1, FTHL17 in GBM to determine whether these antigens can be appropriate target in immunotherapy of GBM.

fifty pathologically confirmed GBM paraffin embedded tissue sample were conducted into this experiment. Total RNA was extracted from these samples and TaqMan based RealTime PCR technology was used to evaluation of HORMAD, ADAM29 and FTHL17 gene expression.

according to our results HORMAD1 is the most frequent expressed gene in these tissue samples. Forty-four percent of samples (22 out of 50 samples) expressed HORMAD1 in various levels. Either FTHL17 and ADAM29 were expressed in only one of samples .On the other hand, according to statistical studies of patients' demographic findings and the expression of genes, HORMAD1 has had a much higher level of expression in glioblastoma samples than other genes, which, based on these results, we can consider this gene as a therapeutic target in immunotherapy

CT46/HORMAD1 is a single-copy gene on chromosome 1q21.3, A review of carcinoma showed that 31% of carcinomas express this antigen in high numbers. In 2006, the gene had a high expression in gastric cance. so far no precise and thorough study has been done on the existence (expression) of these gene in glioblastma samples,
 
HORMAD1 can be a promising target in immunotherapy research targeting GBM.
multiform (GBM, World Health Organization grade IV) is the most common and aggressive

Glioblastoma is the most common primary malignancy of the brain, the prognosis of which is poor. Immunotherapy with cancer/testis (CT) antigens is a novel therapeutic approach for glioblastoma. This study aimed to investigate the expression rate of MAGE-E1, GAGE, and SOX-6 in glioblastoma tumors using the immunohistochemistry (IHC) method.

Expression of MAGE-E1, GAGE, and SOX-6 were determined by IHC in 50 paraffin blocks of glioblastoma. The results were compared between variables including age, gender, tumor location, and Karnofsky performance status (Kps) score. Survival analysis was also performed.

The expression levels of SOX-6, MAGE-E1, and GAGE were 82%, 78%, and 76%, respectively. The relationship between CT antigens and age, gender, and tumor location was not significant, while the association between MAGE-E1 expression and age was statistically significant (p =0.002). High expression levels of SOX-6 and MAGE-E1 were associated with low Kps scores (p =0.034 and p <0.001, respectively). Survival analysis showed that age >40 and Kps score p =0.005 and p =0.018, respectively). Expression of MAGE-E1 and GAGE was negatively associated with overall 2-year survival (p =0.001 and p =0.021, respectively).

The expression of all the three CT antigens, especially MAGE-E1 and SOX-6, was high in patients with glioblastoma. It can be concluded that these markers are ideal targets for immunotherapy in these patients. MAGE-E1 and SOX-6 can be considered as important markers in determining the prognosis of glioblastoma.

Breast cancer is the most prevalent cause of cancer death in women worldwide. The increasing prevalence and high mortality rate of this cancer has made it necessary to understand the molecular mechanism. Axillary lymph node involvement plays a great role in 5 year survival rate. So, identifying specific biomarkers for early detection of lymph node involvement in breast cancer patients can greatly increase survival rates and treatment responses.

In this study, 54 negative sentinel lymph nodes and 106 positive sentinel lymph nodes with metastatic involvement were examined from paraffin samples. Cytokeratin 19 (CK19) was used as a marker of lymph node metastasis and its expression was assessed by Real-time PCR (qPCR) and absolute quantification.

Higher cytokeratin 19 mRNA copy number was identified in metastatic lymph nodes group (P=0.005). The area under the ROC curve (AUC) was significant with P=0.006., the specificity and sensitivity were 63% and 89%, respectively for breast cancer detection on the basis of ROC curve analyses determining the optimum cut-off 250.we demonstrated that CK 19 overexpression was significantly correlated with tumor size, lymph nodes metastasis and pathological stage

Our data indicate that simultaneous use of molecular and conventional diagnostic methods can increase the sensitivity and specificity of pathologic results and decrease the false negative cases.

Food hypersensitivity to walnut usually results in mild symptoms; however, several cases of anaphylactic reactions to this product have been observed. This study aimed to determine the immunochemical characteristics of the Persian walnut and provide the recombinant form of its main allergen. The allergenic proteins of the Persian walnut were extracted by standard procedure. The IgE-binding profile was determined by common immunoassays, including ELISA and Western blotting. The characteristics of the main allergenic protein which showed a stronger and higher frequency of IgE-reactivity with the patient’s sera was identified by MALDI-TOF-TOF method. The conventional PCR was used for the amplification of the coding sequence of the target protein which was then inserted into pET-21b(+) vector and expressed in E. coli BL21. The recombinant allergen was purified by metal affinity chromatography and the ELISA and immunoblotting assays were used for the evaluation of the IgE-binding capacity of the recombinant protein. All patients showed a considerable specific IgE-reactivity to total extract (OD 0.58±0.43 versus 0.047±0.026; P<0.05) in ELISA. Immunoblotting with crude extract indicated considerable IgE-reactivity of the patients’ sera with a 15-kDa allergen which was characterized as 2S albumin by mass spectrometry methods. The refolded walnut recombinant 2S albumin showed considerable IgE-reactivity in ELISA and western blotting with patients’ sera. We demonstrated that the refolding of walnut recombinant 2S albumin could result in the reconstruction of an IgE-reactive allergen with a rather similar immunoreactivity to its natural counterpart. The refolded recombinant protein could be a suitable candidate for diagnostic and therapeutic approaches.

Intra-operative molecular diagnostic assays are currently used for the detection of lymph node metastases. The objective of this study was to find new biomarkers to improve diagnostic accuracy in the detection of metastatic axillary lymph nodes in breast cancer patients.

We applied an absolute quantitative real-time reverse transcription-PCR to quantitate the expression of CK19, KLK11, and CLEC3A mRNAs in 79 FFPE sentinel lymph nodes (SLNs) from 35 breast cancer patients. The CK19 was confirmed as a standard biomarker, and the level of expression of selected new markers, KLK11 and CLEC3A, was evaluated in pathologically negative and positive SLNs by using absolute quantitative real-time PCR.

The overall concordance of the CK19 gene with pathological results was 92.4% (less than 250 copies) in negative SLNs and 85% in positive SLNs (more than 250 copies). The sensitivity and specificity of CK19, which were detected by real-time PCR, was 85% and 46%, respectively. Our results revealed that lower CLEC3A was associated with more lymph node involvement. We could set a cut-off point for CLEC3A with the sensitivity of 78% and specificity of 60%. Also, the mean KLK11 had a statistically significant reverse correlation with tumor grade (p = 0.017). Higher CK19 levels were related to more tumor invasion (p < 0.0001).

Regarding the findings, CLEC3A along with CK19 can be used as a promising marker with high sensitivity and specificity for the detection of metastatic SLN.

Allergic Rhinitis (AR) is an IgE-mediated inflammatory disorder with high morbidity rates. The eitiology of this disease is understood to occur from a complex interaction between genetic and environmental factors. T helper type 2 cells have been shown to have a crucial role in atopic disease due to their production of the cytokines, intelukin (IL)-13 and IL-4, involved in inflammation. Research has shown single nucleotide polymorphisms (SNP) of the IL-13 and IL-4 genes to be associated increased levels of IgE and with allergic diseases such as, allergic rhinitis, asthma, and atopic dermatitis. Specifically, the rs2243250 SNP of IL-4 and the rs20541 SNP of IL-13 have been shown to be associated with AR.

A case-control study was designed to investigate the relationship between the two SNPs rs2243250 and rs20541 with the incidence of AR. The SNPs were examined in patients with AR and healthy controls (86 patients and 86 controls). Blood samples were collected and DNA was extracted to evaluate the SNPs by RFLP-PCR.

Recessive analysis model of the IL-13 gene (GG vs. AA+AG) revealed that the GG genotype was more common in AR patients (P=0.36) )OR=0.8 [81% CI 0.38-1.6]). For the IL-4 gene (TC vs. TT+CC), the TC genotype was more common in AR patients (P = 0.0022)) OR=0.71 [60% CI 1.41-5.02]). Furthermore, in the IL-4 gene, the 590 T>C polymorphism had a significant association with AR. However, no association was found between AR and the IL-13 rs20541 polymorphism.

Our findings suggest that the IL-13 polymorphism (rs20541, Exo 4, G>A, Arg130Gln) and IL-4 polymorphism (rs2243250= C-590T, promoter, T>C) are co-associated with AR and sensitivity to aeroallergens. However, this study used a cohort of AR patients and healthy controls from the northeast of Iran. Given the influence of ethnicity and environment on genetics, further investigation is needed to elucidate the role of SNPs in IL-4 and IL-13 in AR among different populations.

Cancer/Testis Antigens (CTAs) are a subgroup of tumor-associated antigens which are expressed normally in germ line cells and trophoblast, and aberrantly in a variety of malignancies. One of the most important CTAs is Developmental Pluripotency Associated-2(DPPA2) with unknown biological function. Considering the importance of DPPA2 in developmental events and cancer, preparing a suitable platform to analyze DPPA2 roles in the cells seems to be necessary.
In this study, the coding sequence of DPPA2 gene was amplified and cloned into the retroviral expression vector to produce recombinant retrovirus. The viral particles were transducted to Esophageal Squamous Cell Carcinoma (ESCC) cell line (KYSE-30 cells) and the stable transducted cells were confirmed for ectopic expression of DPPA2 gene by real-time PCR.
According to the critical characteristics of retroviral expression system such as stable and long time expression of interested gene and also being safe due to deletion of retroviral pathogenic genes, this system was used to induce expression of DPPA2 gene and a valuable platform to analyze its biological function was prepared. Transduction results clearly showed efficient overexpression of the gene in target cells in protein level due to high level of GFP expression.
Such strategies can be used to produce high levels of desired protein in target cells as a therapeutic target. The produced recombinant cells may present a valuable platform to analyze the effect of DPPA2 ectopic expression in target cells. Moreover, the introduction of its potential capacity into the mouse model to evaluate the tumorigenesis of these cancer cells in vivo leads to an understanding of the biological importance of DPPA2 in tumorigenesis. In addition, our purified protein can be used in a mouse model to produce specific antibody developing a reliable detection of DPPA2 existence in any biological fluid through ELISA system.

Human Cripto-1, a member of the EGF-CFC family, is involved in embryonic development, embryonic stem cell maintenance, and tumor progression. It also participates in multiple cell signaling pathways including Wnt, Notch, and TGF-β. Remarkably, it is expressed in cancer stem cell (CSC) compartments, boosting tumor cell migration, invasion, and angiogenesis. Although Cripto-1 is overexpressed in a variety of human malignant tumors, its expression in esophageal squamous cell carcinoma (ESCC) remains unclear. Our aim in this study was to evaluate the possible oncogenic role of Cripto-1 in ESCC progression and elucidate its association with clinicopathological parameters in patients.
In this study, Cripto-1 expression in 50 ESCC tissue samples was analyzed and compared to corresponding margin-normal esophageal tissues using quantitative real-time PCR.
Cripto-1 was overexpressed in nearly 40% of ESCC samples compared with normal tissue samples. Significant correlations were observed between Cripto-1 expression and tumor differentiation grade, progression stage, and location (p Our results indicate that overexpression of Cripto-1 is involved in the development of ESCC. Further assessment will be necessary to determine the role of Cripto-1 cross talk in ESCC tumorigenesis.

The development of allergic rhinitis (AR) is caused by the interaction between genetic predisposition and environmental factors. In this study, the association between GATA3 single nucleotide polymorphisms and AR in an Iranian population was identified.
This case-control study was performed on 86 patients with AR and 86 healthy subjects. This study aimed to evaluate a potential association between two GATA3 SNPs, rs1269486 and rs2229360, and AR. Blood samples were collected and DNA was extracted for the evaluation of these SNPs by RFLP-PCR.
A statistically-significant association was found between rs1269486 and AR (P SNP rs1269486 of GATA3 was associated with AR and sensitivity to aeroallergens in our population. Because of the significance of this gene in AR, studying the association between GATA3 polymorphisms and AR is recommended for other populations.

Variation in microsatellite sequences that are dispersed in the genome has been linked to a deficiency in cellular mismatch repair system and defects in several genes of this system are involved in carcinogenesis. Our aim in this study was to illustrate microsatellite DNA alteration in esophageal cancer.
DNA was extracted from formalin fixed paraffin embedded (FFPE) tissues from surgical and matched margin-normal samples. Microsatellite instability (MSI) and loss of heterozygosity (LOH) were studied in 50 cases of esophageal squamous cell carcinoma (ESCC) by amplifying six microsatellite markers: D13S260 (13q12.3), D13S267 (13q12.3), D9S171 (9p21), D2S123 (2p), D5S2501 (5q21) and TP53 (17p13.1) analyzed on 6% denaturing polyacrylamide gel electrophoresis.
Statistical analysis indicated a near significant reverse correlation between grade and LOH (P= 0.068, correlation coefficient= -0.272). Specifically, increased LOH in tumor DNA has a significant correlation with increased differentiation from poorly differentiated to well differentiated tumors (P= 0.002 and P= 0.016 respectively). In addition, higher number of chromosomal loci with LOH showed a reverse correlation with lymph node metastasis (P= 0.026, correlation coefficient= -0.485). Furthermore, there was a positive correlation between addiction and MSI (P= 0.026, correlation coefficient= 0.465).
Microsatellite DNA alterations may be a prognostic tool for detection and the evolution of prognosis in patients with SCC of esophagus. It can be concluded that regional lymph node metastasis would be less likely with increased heterozygote loci and addiction with any of opium, cigarette, water pipe or alcohol can be a susceptibility factor(s) for MSI.

Sublingual allergen-specific immunotherapy is a safe and effective method for treatment of IgE-mediated respiratory allergies; however, the underlying mechanisms are not fully understood. This study was planned to test whether sublingual immunotherapy (SLIT) can exert epigenetic mechanisms through which the airway allergic responses can be extinguished. BALB/c mice were sensitized intraperitoneally and challenged intranasally. Then, they received sublingual treatment with recombinant Che a 2 (rChe a 2), a major allergen of Chenopodium album. After SLIT, allergen-specific antibodies in sera, cytokine profiles of spleen cell cultures, mRNA and protein expression of lung-derived IL-33, IL-25, and TSLP (thymic stromal lymphopoietin), and histone modifications of these three genes were assessed. Following Immunotherapy, systemic immune responses shifted from Th2 to Th1 profile as demonstrated by significant decrease in IgE and IL-4 and substantial increase in IgG2a and IFN-γ. At local site, mRNA and protein levels of lung-derived pro-inflammatory cytokines IL-33 and TSLP were markedly down-regulated following SLIT that was associated with marked enrichment of trimethylated lysine 27 of histone H3 at promoter regions of these two cytokines. In our study, sublingual immunotherapy with recombinant allergen effectively attenuated allergic immune responses, at least partly, by induction of distinct histone modifications at specific loci. Additionally, the lung-derived pro-allergic cytokines IL-33 and TSLP could be promising mucosal candidates for either monitoring allergic conditions or therapeutic approaches.

Recently, it has been revealed that tyrosine kinase inhibitors (TKIs) act through inducing both oxidative and endoplasmic reticulum (ER) stress in chronic myeloid leukemia cells. However, ER stress signaling triggers both apoptotic and survival processes within cells. Nevertheless, mechanisms by which TKIs avoid the pro-survival effects are not clear. The aim of this study was to evaluate the potential role of oxidative stress in activity of unfolded protein response (UPR) survival pathway within K562 cell line. The expression of UPR survival target genes, Xbp1, and Grp94 (glucose requiring protein 94) was studied in single and combined exposure to oxidative and ER stress in K562 cell line by quantitative and qualitative PCR. The expression of UPR-related survival gene Grp94 was hampered by exposing to oxidative stress in cell induced with ER stress. Interaction of oxidative and ER stress may role as a mediator influencing UPR signaling activity.

Aim and Cancer/testis (CT) antigens are a category of tumor antigens that are typically expressed only in the human germ line, and in several types of tumor. MAGEA4 is one member cancer testis antigen family that has relation with other tumors. Since the biological function of MAGEA4 is unclear, the aim of the present study was amplification and cloning of the gene in the expression vector to produce recombinant protein that expresses MAGEA4. Using PCR specific primers including restriction site, MAGEA4 was amplified. The purified PCR products were ligated between the BamH1 and Xho1 sites of pTZ57/R cloning vector and transformed into Escherichia coli Top10 strain and screened by IPTG and X-Gal. The correct orientation of MAGEA4 fragment was identified by restriction enzyme analysis and sequencing of constructed plasmid. Then sub cloning was carried out within pRUF expression vector with the same restriction site. The final confirmation was performed using colony PCR, double digesting and sequence analysis. Therefore the MAGEA4 gene was cloned into the restriction site of pRUF. This study is an important step for producing recombinant proteins and is used to find the function of this gene for therapeutic targets.

CD44 is a member of the cell adhesion molecules family. Naturally, CD44S, along with CD44V3 influence the cell motility, migration, and adhesion, while in tumor cells they lead to tumor invasion, progression, and metastasis. The purpose of this research is to evaluate the CD44S and CD44V3 expression in Esophageal Squamous Cell Carcinoma (ESCC) and to reveal their correlations with clinicopathological features of patients. Fresh tumoral and distant tumor-free esophageal tissues were obtained from 50 patients with ESCC. Using quantitative real-time PCR, the expression levels of CD44S and CD44V3 were quantified and compared in both groups of cells. The patients had not received any therapeutic interference, such as chemotherapy or radiation, prior to sampling. Significant overexpression of CD44S and CD44V3 mRNA was observed in 13 (26.0%, P=0.03) and 11 (22.0%, P=0.007) tumor specimens, respectively. The expression of the genes were significantly correlated not only with each other (P=0.0001), but also with differentiation grade of tumor (P=0.033), stage of tumor progression (P=0.003), and depth of tumor invasion (P=0.00). In addition, low level of CD44V3 mRNA expression was attended to be associated with tumor invasion. There is no correlation between CD44S expression with clinicopathological features of patients; however, simultaneous expression of these genes has an important effect on tumorigenesis.

Cancer/testis antigens (CTAs) are a subgroup of tumor-associated antigens which are expressed in trophoblast، ovary and testis germ line cells. CTAs are also expressed in a variety of cancer cells and their biological function in germ line and tumors cells is unclear yet. One of the most important CTAs is CT100 which is also known as DPPA2. The aim of this study was isolation، amplification and cloning of DPPA2 coding sequence. Having examined the gene expression pattern of different cell lines، we amplified coding sequence of DPPA2 gene and cloned it in pTZ57R/T vector. Recombinant colonies were analyzed and confirmed by colony PCR، double digestion and sequencing. Considering the importance of DPPA2 roles in the development، its role analysis in carcinogenesis seems valuable. Our results prepare a suitable platform to analyze its biological function.

Asthma results from the interaction between genetic and environmental factors. ADAM33 gene on chromosome 20p13 is associated with asthma and airway hyperresponsiveness. This is a case-control study, where four SNPs S1 (rs3918396), T1 (rs2280091), T2 (rs2280090), V4 (rs2787094) of ADAM33 gene have been assessed in patients with allergic asthma and normal controls (95 patients and 86 normal). Blood samples of these participants have been genotyped by PCR and the RFLP method. There was no association between asthmatic patients and polymorphisms of alleles, genotypes and haplotypes of the ADAM33 gene. When categorizing the asthmatic patients in severe, moderate and mild groups, associations in the subcategories of asthmatic patients were found. There were associations between polymorphisms of C allele of T1 SNP with severe asthmatic patients and G allele of V4 SNP with moderate asthmatics respectively (P=0.006, P=0.01). There was a significant association between sensitivity to mite and polymorphism of C allele of T1 SNP (P=0.02). Besides, there was a significant association between sensitivity to weeds and genotype GG of V4 SNP (P=0.05). Polymorphisms of ADAM33 gene might be associated with severe asthma and sensitivity to aeroallergens in northeast of Iran, but further studies are needed to determine the polymorphisms in this area and other regions of our country.

CatSper genes are a novel family of four sperm-specific calcium channels, which indicate testis-specific expression patterns. Despite the crucial role of CatSper genes in the male reproduction, very little is known about the factors that regulate their expression. The objective of this study was to investigate the effects of vitamin E treatment on the expression of CatSper 1 and CatSper 2 genes as well as sperm quality in the aged male mice. Twenty four 11-12 months old aged male mice and twenty four 2-3-months old young male mice were randomly divided into four groups. Control groups received no injection. The experimental groups of male mice were received intraperitoneal injection of 106 mg/kg vitamin E daily for 35 days. Left testis and cauda epididymides from each mouse were collected on the days 21, 28 and 35 following vitamin E treatment and were used for Real-Time PCR and immunohistochemistry. Also, sperm analysis was performed according to the WHO guidelines given for human sperm examination. Data were analyzed using SPSS software. Administration of vitamin E improved sperm parameters in the aged as well as young adult male mice. In addition, the expression of CatSper genes increased following vitamin E treatment. Also, intensity of signal for CatSper1 and CatSper2 increased in the head and middle piece of sperm in experimental group as compared to those of control ones. The vitamin E treatment significantly improved the sperm quality, especially in terms of sperm motility, count and morphology rate. Furthermore, CatSper genes expression could be up-regulated by the vitamin E treatment.

This study was conducted to optimize a highly efficient mRNA transfection into dendritic cells (DC) derived from esophageal squamous cell carcinoma (ESCC) patients. Applying an electroporation technique, in vitro synthesized Green Fluorescent Protein (GFP) mRNA was transfected as an indicator into the DCs derived from a healthy donor. Flow cytometry revealed 84.9% transfection efficiency for DCs transfected with GFP mRNA. Optimized condition (500V/300 ms) yielded 79.8% efficiency in transfecting GFP mRNA into DCs from ESCC patient. Applying this efficient method, tumoral mRNA was transfected into DCs. T cells were then primed with tumor RNA/DCs and cytotoxicity assay revealed significantly higher lyses of tumor/DC vs. Mock DC; approving the optimized method for further establishment of the preclinical phase of DC-based immunotherapy for ESCC.Keywords: Immuno-gene therapy; Esophageal squamous cell carcinoma (ESCC); Electroporation; mRNA transfection; Dendritic cell; GFP