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عضویت
فهرست مطالب نویسنده:

mohammad khalaj-kondori

  • Safa Jasim Tuama Ali, Mohammad Khalaj-Kondori*, Mohammadali Hoseinpour Feizi, Mehdi Haghi
    Background

    People with autism frequently exhibit poor social skills, communication difficulties, and repetitive and stereotyped behaviors. MicroRNAs (miRNAs) are potential and promised targets in developing of new treatment strategies for autism.This study aimed to assess the relative expression of miR-124a, miR-34a-3p, miR-545-3p, miR-153, and BDNF in the blood samples of autistic children.

    Methods

    The children autism rating scale (CARS) was used to determine the severity of autism and to confirm the diagnosis. Blood samples were obtained from 50 patients and 40 age-/sex-matched healthy controls. Expressions of miR-545-3p, miR-34a-3p, miR-124a, and BDNF were evaluated using qRT-PCR. Pearson's correlation coefficient and regression analysis were used to check correlations between relative expressions of the miRNAs and BDNF. Biomarker potencies were assessed by ROC curve analysis.

    Results

     qRT-PCR analysis showed that the relative expressions of miR-545-3p, miR-34a-3p, miR-124a, and BDNF were significantly higher in the patients' group than the healthy controls. However, the relative expression of miR-153 was significantly lower in the case group than the control group. The relative expression of miR-124a was positively correlated with those of miR-545-3p and BDNF among the patients group. Also, the relative expressions of miR-545-3p and BDNF were positively correlated with each other. The ROC curve data also indicated that miR-124a, miR-34a-3p, miR-545-3p, miR-153, and BDNF could be possible diagnostic biomarker for CARS diagnosis (AUC=0.8328, AUC=0.8354, AUC=0.6727, AUC=0.8518 and AUC=0.8214, respectively).

    Conclusions

    Deregulation of miR-124a, miR-454-3p and BDNF might be considered as potential biomarkers for severity of autism.

    Keywords: Autistic Children, Autism, Biomarker, Brain-Derived Neurotrophic Factor, Gene Expression, Mirna
  • Rawan Hassan Al-Saeedi, Mohammad Khalaj-Kondori*, Mohammadali Hosseinpour Feizi, Jafar Hajavi
    Background

    Inflammation contributes to cancer pathobiology through different mechanisms. Higher levels of pro-inflammatory cytokines can lead to hyperinflammation and promote cancer development and metastasis. For cancer treatment, Doxorubicin (DOX) can be encapsulated into the poly-lactic-glycolic acid (PLGA) nanoparticles. This study aimed to investigate the impact of doxorubicin-loaded PLGA nanoparticles (DOX-PLGA NP) on the expression of pro-inflammatory genes TNF-α, IL-6, iNOS, and IL-1β in the MCF-7 cells.

    Methods

    The DOX-PLGA NP was prepared by loading doxorubicin into PLGA and characterized using dynamic light scattering (DLS) and atomic force microscopy (AFM). The cytotoxic effect of the nanoparticles was determined by the MTT assay, and their impacts on the expression of pro-inflammatory genes were assessed by qRT-PCR.

    Results

    The encapsulation efficiency and loading capacity were 60±1.5 and 1.13±0.21 percent, respectively. The zeta potential and mean DOX-PLGA nanoparticle size were -18±0.550 mV and 172±55.6 nm, respectively. The 50% inhibitory concentration (IC50) of the DOX-PLGA NP on MCF-7 cell viability was 24.55 µg/mL after 72 hours of treatment. The qRT-PCR results revealed that the 20 µg/mL concentration of the DOX-PLGA NP significantly suppressed the expression of the pro-inflammatory genes TNF-α, IL-6, iNOS, and IL-1β compared to DOX alone (20 µg/mL). Additionally, the suppression effect of DOX-PLGA NP on the expression of these pro-inflammatory genes was dose-dependent.

    Conclusion

    These results show that DOX-PLGA NP efficiently suppressed the expression of pro-inflammatory genes. Furthermore, encapsulation of DOX into PLGA nanoparticles significantly improved the effectiveness of DOX in suppressing pro-inflammatory genes in MCF-7 breast cancer cells.

    Keywords: Breast Cancer, Cytokines, Doxorubicin, Polylactic Acid-Polyglycolic Acid Copolymer, Pro-Inflammatory Cytokine
  • Basheer Kadhum Kharmeet, Mohammad Khalaj-Kondori*, Mohammadali Hosseinpour Feizi, Jafar Hajavi
    Background

    Pro-inflammatory cytokines play critical roles in cancer pathobiology and have been considered potential targets for cancer management and therapy. Understanding the impact of cancer therapeutics such as 5-fluorouracil (5-FU) on their expression might shed light on development of novel combinational therapies. This study aimed to  encapsulate 5-FU into PLGA  and evaluate their effects on the expression of pro-inflammatory genes IL-9, IL-17-A, IL-23, and IFN-γ in the HT-29 cells.

    Methods

    PLGA-5-FU NPs were constructed and characterized by Dynamic Light Scattering (DLS) and Atomic Force Microscopy (AFM). The cytotoxicity was evaluated by MTT test and, the IC50 was identified. HT-29 cells were treated with different concentrations of the PLGA-5-FU NPs for 48 hours and, gene expression levels were analyzed by qRT-PCR.

    Results

    DLS and AFM analysis revealed that the prepared PLGA-5-FU NPs were negatively charged spherical-shaped particles with a mean size of 215.9 ± 43.3 nm. PLGA-5-FU NPs impacted the viability of HT-29 cells in a dose- and time-dependent manner. The qRT-PCR results revealed a dose-dependent decrease in the expression of IL-9, IL-17A, IL-23 and IFN-γ genes, and their expressions were significantly different in both 10 and 20 µg/mL treated groups compared to the control. However, although the treatment of HT-29 cells with 20 µg/mL free 5-FU resulted in decreased expression of the studied genes, the differences were not statistically significant compared to the control group.

    Conclusions

    PLGA-5-FU NPs significantly suppressed expression of the IL-9, IL-17A, IL-23 and IFN-γ genes, and the encapsulation of 5-FU into PLGA improved considerably impact of the 5-FU on the HT-29 cells.

    Keywords: Cancer Therapy, Colorectal Cancer, Fluorouracil, Polylactic Acid-Polyglycolic Acid Copolymer (PLGA), Pro-Inflammatory Cytokine
  • Zeynab Shaghaghi Torkdari, Mohammad Khalaj-Kondori, MohammadAli Hosseinpour Feizi
    Background

    Breast cancer is identified as the most common malignancy and cause of cancer-related death worldwide. Compared with healthy controls, this study evaluated the expression level and diagnostic power of lncRNA plasma TINCR in breast cancer patients.

    Materials and Methods

    Fifty-eight women diagnosed with invasive ductal carcinoma and fifty healthy age-matched controls were included in the study. TRIzol® LS regent was used to isolate the total RNA from the whole plasma. Total RNA was converted to cDNA using Prime ScriptTM RT reagent kit and the expression levels of TINCR were quantified by qRT-PCR.

    Results

    Low levels of TINCR lncRNA were observed in the plasma of breast cancer patients compared with control subjects. Plasma TINCR level was also positively correlated with the diagnostic age of breast cancer patients.

    Conclusion

    A low level of plasma TINCR could discriminate breast cancer patients from healthy control subjects.

    Keywords: Breast cancer, TINCR, Plasma
  • Amin Moqadami, Alireza Ahmadi, Mohammad Khalaj-Kondori *
    Background

    Colorectal cancer (CRC) is the third mostcommoncancer that frequently spreads to other parts of thebody, with a low chance of recovery and a high mortality rate. Long non-coding RNAsare considered significant prognostic and diagnostic indicators because they are dysregulated in various cancers and have distinctive expression patterns and high tissue- and cell-specificity. VLDLR-AS1 lncRNA deregulation has been associated with several malignancies.

    Objectives

    This study aimed to evaluate the expression levels of VLDLR-AS1 in CRC. It was the first time this assessment was conducted.

    Methods

    We studied 188 samples, including 94 tumor samples and 94 paired adjacent non-tumor tissues. Total RNA was extracted, and its quantity and purity were assessed. TaKaRa PrimeScript 1st Strand cDNA Synthesis Kit (Kusatsu, Japan) was used for the reaction. The StepOnePlus Real-Time PCR System (Applied Biosystems) was set up to assess the relative expression of VLDLR-AS1.

    Results

    According to the qRT-PCR data, the VLDLR-AS1 expression levels in CRC tissues were significantly lower than in tumor margins (P < 0.0001). In addition, receiver operating characteristic curve analysis demonstrated that VLDLR-AS1 expression can discriminate between tumor and non-tumor samples with the sensitivity and specificity of 72.34% and 51.06%, respectively (P = 0.03, AUC = 0.6274). However, no significant association was found between the expression levels of VLDLR-AS1 and clinicopathological features in CRC patients.

    Conclusions

    The results of this study indicated that VLDLR-AS1 lncRNA is significantly downregulated in the tumor tissues of CRC patients compared to healthy tumor margin tissues. This evidence shows the potential of this gene as a promising biomarker for the early detection of CRC development.

    Keywords: Colorectal Cancer, LncRNA, VLDLR-AS1, Biomarker
  • Arezu Karimpur Zahmatkesh, Amin Moqadami, Mohammad Khalaj-Kondori *

    Radiotherapy (RT) has been commonly applied to treat advanced local cancers. In radiation therapy, high doses of radiation are utilized to trigger cell death. Radiation often leads to DNA double-strand breakages (DSB), which causes the activation of downstream genes including those for non-coding RNAs (ncRNA) such as long non-coding and RNAsmicro RNAs. The consequence of RT significantly relies on the radiosensitivity of cancer cells, which is affected by multiple factors, including some proteins and cellular processes. Activation of these genes can cause cell cytotoxicity and indirectly damages the cells. Recent studies have shown that non-coding RNAs can play as radiosensitivity or radioinhibitory regulators in cancers by mechanisms such as cell cycle arrest or affecting the DNA damage repair systems. ncRNAs are also known to function as tumor suppressor genes or oncogenes in colorectal cancer and therefore are considered potential diagnostic biomarkers in disease detection. For example, the investigations have shown that miR-29a and miR-224 can be informative biomarkers for early detection or screening of CRC via a noninvasive method such as liquid biopsy. Here, we discuss ncRNAs involved in the radioresistance and radiosensitivity of CRC and highlight their predictive clinical value in response to RT. Accordingly, this review represents a principal guide in the context of three major types of ncRNAs with potential roles in the pathway of radiosensitivity and radioresistance, including miRNAs, lncRNAs, and circRNAs which can be considered a precious archivement in organizing additional studies and broadening views in this area. Our findings can also assist radiotherapists in predicting CRC patients’ response and, therefore, prognosis to radiation therapy, although, to achieve our goals in the clinic, we certainly need further studies.

    Keywords: Colorectal cancer, Non-coding RNAs, Radioresistance, Radiosensitivity Radiotherapy
  • Amin Moqadami, Mohammad Khalaj-Kondori *
    Long non-coding RNAs (lncRNAs) have recently emerged as effective regulatory agents in biological processes as well as in the formation of tumors. LncRNAs are important regulators of cell transformation and cancer progression. LncRNA NEAT1 is one of the most important lncRNAs, and its deregulation has been reported in a variety of human cancers. Ovarian cancer has an inverse relationship with the number of reported pregnancies and deliveries, while it has a direct relationship with infertility. This study aimed to investigate NEAT1 expression in ovarian cancer. A total of 140 tissue samples, including 70 ovarian tumors and 70 marginal samples, were included in the study. Total RNA was extracted using the RNXplus solution. The quality and quantity of the extracted RNAs were determined using gel electrophoresis and a NanoDrop device. The complementary DNA was synthesized by the reverse transcriptase enzyme, and quantitative reverse transcriptase PCR was used to quantify the expression of NEAT1. A comparison between the mean expression of NEAT1 in ovarian tumors and marginal samples showed an increase in NEAT1 expression in tumor tissue that was not statistically significant (P-value = 0.2). ROC curve analysis also showed that NEAT1 expression level might not be an informative biomarker for ovarian cancer.
    Keywords: NEAT1, ovarian cancer, qRT-PCR, Biomarker
  • بهارک ابراهیمی بهنام، محمد خلج کندری*، محمدعلی حسینپور فیضی
    زمینه و هدف

    سقط مکرر (RSA) شایع ترین عارضه بارداری است که به وقوع دو یا بیش از دو مورد سقط قبل از هفته بیستم بارداری اطلاق می گردد. یکی از عوامل مهم دفع جنین مولکول ایمونوتولرانسHLA-G است که نقش مهمی در حفاظت از جنین در قبال سیستم ایمنی مادر دارد. هدف از این مطالعه بررسی ارتباط پلی مورفیسم های rs1632943 و rs1736932 ناحیه پروموتر ژن HLA-G با سقط مکرر در زنان شمال غرب ایران است.

    مواد و روش ها

     در این مطالعه مورد- شاهدی، 100 زن با سابقه سقط مکرر به عنوان گروه مورد و 80 زن سالم با یک یا بیشتر از یک کودک به عنوان گروه کنترل انتخاب شدند. DNA ژنومی از نمونه خون محیطی آن ها استخراج و ژنوتیپ های آن ها با استفاده از روش تعیین توالی مشخص شد. بررسی آماری نتایج حاصل، توسط آزمون مربع کای و نرم افزار SPSS ورژن 16 انجام شد.

    یافته ها

    در پلی مورفیسم rs1632943 فراوانی ژنوتیپ های CC، CA و AA در گروه بیمار به ترتیب 8 ٪، 33 ٪و 59 ٪و در گروه شاهد 25/16٪، 75/43٪ و40٪ بود. تجزیه و تحلیل آماری نشان داد که ژنوتیپ AA با سقط مکرر خود به خود همراه است (p=0.005) و در پلی مورفیسم rs1736932 فراوانی ژنو تیپ هایCC، CG و GG به ترتیب 8٪، 32٪و 60٪ در گروه بیمار و 5/17٪، 25/41٪ و 25/41٪ در گروه شاهد بود. تجزیه وتحلیل آماری نشان داد که ژنوتیپ GG با سقط مکرر همراه است (005/0 P=). همچنین بررسی هاپلوتایپی نشان داد که هاپلوتایپ H1 (GA) با بیماری همراهی نشان می دهد.

    نتیجه گیری

    نتایج مطالعه حاضر حاکی از آن است که پلی مورفیسم های rs1632943 و rs1736932 می توانند به عنوان یک عامل خطر برای RSA در زنان شمال غربی ایران در نظر گرفته شود.

    کلید واژگان: آنتی ژنهای HLA-G, سقط مکرر, پلی مورفیسم تک نوکلئوتیدی, مطالعات همراهی ژنتیکی
    Baharak Ebrahimi Behnam, Mohammad Khalaj Kondori*, Mohammad Ali Hoseinpour Feizi
    Background and Aim

    Recurrent spontaneous abortion (RSA) is the most common complication of pregnancy that refers to two or more miscarriages before the 20th week of pregnancy. HLA-G immunoglobulin molecule plays an important role in protecting the fetus against mother's immune system. The aim of this study was to investigate the association between rs1632943 and rs1736932 polymorphisms with recurrent spontaneous abortion in Northwest of Iran.

    Materials and Methods

    This case-control study included 100 women with history of RSA as our case group and 80 healthy women with one or more than one children as the control group. Genomic DNA was purified from their peripheral blood samples and their genotypes were determined by PCR-sequencing method. Using SPSS 16, statistical analysis was performed by chi-square test.

    Results

    In rs1632943 polymorphism the frequency of CC, CA and AA genotypes were 8%, 33% and 59% in the patient group and 16.25%, 43.75% and % 40 in the control group, respectively. Statistical analysis showed that AA genotype was associated with the recurrent spontaneous abortion (P = 0.005). In the rs1736932 polymorphism, the frequency of CC, CG and GG genotypes were 8%, 32% and 60% in the patient group and 17.5%, 41.25% and 41.25% in the control group, respectively. Statistical analysis showed that GG genotype was associated with the recurrent miscarriage (P = 0.005). Also, haplotype analysis showed that H1 haplotype (GA) is associated with the disorder.

    Conclusion

    Results of the study showed that rs1632943 and ra1736932 polymorphisms

    Keywords: HLA-G antigens, recurrent miscarriage, Single nucleotide polymorphism, Genetic association studies
  • زینب شقاقی ترکداری، محمد خلج کندری*، محمدعلی حسین پور فیضی
    مقدمه

    سرطان پستان مهم‏ترین علت مرگ و میر ناشی از سرطان در زنان است که بررسی عوامل ایجاد کننده یا تشدید کننده آن ضروری به نظر می‏ رسد. در این مطالعه، سطح  mRNA ژن PHB2 در بافت تومور و بافت غیرتوموری مجاور مربوط به 50 زن مبتلا به کارسینوم داکتال غیرتهاجمی پستان مورد بررسی قرار گرفت.

    روش بررسی

    محلول RNX-Puls برای استخراج RNA کل از بافت توموری و غیرتوموری مجاور بیماران مبتلا به سرطان پستان استفاده شد. سپس با استفاده از کیتPrimeScriptTM RT، RNA کل به cDNA تبدیل شد. سطح mRNA ژن PHB2 توسط qRT-PCR مورد بررسی کمی قرار گرفته و داده ‏ها با آزمون تی زوجی مورد آنالیز قرار گرفت. علاوه بر این، بررسی منحنی ROC برای ارزیابی پتانسیل بیومارکری PHB2 در بافت‏ های توموری سرطان پستان انجام شد.

    یافته ‏ها

    سطح پایینی از PHB2 mRNA در بافت توموری بیماران مبتلا به سرطان پستان در مقایسه با بافت غیرتوموری مجاور مشاهده شد (P<0.0001). به عبارت دیگر، سطح PHB2 mRNA  در بافت توموری نسبت به حالت نرمال تقریبا 5/2 برابر کاهش داشته است (P=0.001). آنالیز منحنی ROC نشان داد که سطح mRNA ژن PHB2 می‏ تواند بافت‏ های توموری و غیرتوموری را با اختصاصیت 3/91% و حساسیت 3/63% تشخیص دهد (AUC=0.712, P<0.001).

    نتیجه‏ گیری

    کاهش سطح mRNA ژن PHB2 در بافت توموری بیماران مبتلا به سرطان پستان نشان می‏ دهد که سطح mRNA این ژن احتمالا بافت توموری را از بافت ‏های غیرتوموری مجاور می‏ تواند تمایز دهد.

    کلید واژگان: سرطان پستان, PHB2, کارسینومای داکتال, بیومارکر
    Zeynab Shaghaghi Torkdari, Mohammad Khalaj-Kondori*, Mohamad Ali Hosseinpour
    Introduction

    Breast cancer is the most important cause of cancer mortality among women, therefore the study of its causative or aggravating factors seems necessary. In this study, the mRNA levels of the PHB2 gene were evaluated in tumor and adjacent non-tumor tissues of 50 women diagnosed with invasive ductal carcinoma of the breast.

    Methods

    RNX-Plus solution was used to isolate total RNA from tumor and adjacent non-tumor tissues of breast cancer patients. Thereafter, total RNA was converted to cDNA using PrimeScriptTM RT reagent kit. The mRNA levels of PHB2 were quantified by qRT-PCR and data were analyzed with a paired-samples t test. Furthermore, ROC curve analysis was performed to evaluate the diagnostic capacity of PHB2 expression in breast cancer tumor tissues.

    Results

    A significantly lower level of PHB2 mRNA was observed in tumor tissues of breast cancer patients compared with adjacent non-tumor tissues (P<0.0001). PHB2 mRNA levels showed an approximately 2.5-fold reduction in tumor tissues compared with normal tissues (P = 0.001). Roc cure analysis showed that PHB2 mRNA level can discriminate tumors from non-tumor tissues with 91.3% specificity and 64.3% sensitivity (AUC=0.712, P<0.001).

    Conclusion

    Downregulation of PHB2 in breast cancer patients shows that its mRNA levels can possibly discriminate tumors from non-tumor tissues.

    Keywords: Breast Cancer, PHB2, Ductal Carcinoma, Biomarker
  • Mohammad Khalaj Kondori *, Mina Khajehdehi, Zeinab Shaghaghi Torkdari

    Alzheimer's disease (AD) is a multifactorial disorder that its progress and development are related to various genetic and environmental factors. The disease onset is affected by both genetic and environmental factors such as oxidative stress, inflammation, and mitochondrial dysfunction, and is remarkably related to age progress. Aluminum, a neurotoxic environmental factor, impairs memory performance and can cause neurodegenerative diseases such as AD. On the other hand, the regulatory RNA-binding product of Fragile X mental retardation (FMR1) gene exerts a translational inhibitory effect on the expression of amyloid precursor protein (APP), the main culprit in AD development. In the present study, we treated AlCl3-induced Alzheimer’s disease model rats with Frankincense and investigated its protective and therapeutic effects on the AlCl3-induced memory disturbance by behavioral and molecular assays. Also, Rivastigmine was used as a standard control. Morris Water Maze (MWM) was used to assay special memory working of the rats and quantitative real-time PCR (qRT-PCR) was applied to investigate the expression profile of the FMR1 gene in the hippocampus of the treated rats. MWM behavioral tests indicated that both Frankincense and Rivastigmine not only may prevent AlCl3-induced memory impairment but also may alleviate the memory declines induced by AlCl3 in the rats. Expression analysis showed significant upregulation of the FMR1 gene in response to both Frankincense and Rivastigmin treatments. Further, qRT-PCR results revealed that the AlCl3-induced downregulation of the FMR1 gene expression could significantly be reversed by both Frankincense and Rivastigmine, though Rivastigmine was more effective than Frankincense. In conclusion, our results highlighted that Frankincense might be effective both in the prevention and treatment of memory impairments, to some extent, by affecting the FMR1 gene expression.

    Keywords: Alzheimer's disease, Frankincense, Morris Water Maze, Rivastigmine
  • Zahra Najafi, Mohammad Khalaj Kondori*, MohammadAli Hosseinpour Feizi, Shamsi Abbasaliizadeh
    Background

     Recurrent Pregnancy Loss (RPL) is a multifactorial disease that affects 1-3% of couples. Since Human Leukocyte Antigen-G (HLA-G) gene is involved in fetal maternal immune tolerance, mutations in the HLA-G gene can affect the success rate of pregnancy.

    Objective

     The present study aims to investigate the haplotype effect of rs1736933 and rs2735022 polymorphisms found in the HLA-G gene on the RPL.Methods In this case-control study, participants were 100 women with RPL and 80 women with normal fertility in northwestern Iran. The HLA-G gene promoter was amplified by Polymerase Chain Reaction (PCR) method and sequenced. The genotype and allele frequencies of the two polymorphisms were compared between the two groups by using t-test in SPSS software version 22. Haplotype analysis was performed using PHASE 2.1 and Haploview 4.2 applications

    Findings

     C allele and CC genotype in rs2735022 polymorphism and the G allele and GG genotype in rs1736933 polymorphism showed a significant association with the RPL (P<0.05). Frequency of haplotypes AA, AC, GA, GC were 0.72, 0.23, 0.01, 0.03 in patients and 0.39, 0.01, 0.02, 0.59 in the control group (P<0.05). The linkage disequilibrium score was 94.

    Conclusion

     Analysis of GC haplotype in rs2735022 and rs1736933 polymorphisms of the HLA-G gene can be helpful in genetic studies of women with RPL.

    Keywords: Recurrent pregnancy loss, Polymorphism, Haplotype, Human Leukocyte antigen-G
  • Seyedeh Nahid Fotuhi, Mohammad Khalaj-Kondori *, Hadis Karimi

    Patients with ovarian cancer are mostly diagnosed at advanced stages which leads to poor prognosis and high mortality rate. Deregulation of lncRNA HOXD-AS1 expression associates with cancer development and metastasis. However, the expression level of this lncRNA in ovarian cancer is not determined.50paired ovarian tumors and their adjusted normal tissues were included in the study. Total RNA was extracted by TRIzol® Reagent and reverse-transcribed to cDNA using PrimeScript II cDNA synthesis kit. The expression levels of HOXD-AS1 were quantified by qRT-PCR and compared. The Roc curve analysis was used to evaluate the capacity of HOXD-AS1 as a biomarker for ovarian cancer. We observed that lncRNA HOXD-AS1 was significantly upregulated in ovarian tumors compared to their adjusted normal tissues (p <0.003). Moreover, the ROC curve analysis revealed that the lncRNA HOXD-AS1 expression level could discriminate tumoral and non-tumoral tissues with 85% sensitivity and 88% specificity. The lncRNA HOXD-AS1 expression level might be considered as a potential biomarker for ovarian cancer development.

    Keywords: ovarian cancer, Biomarker, lncRNA, HOXD-AS1, Gene expression
  • لیلی خاصوانی، محمد خلج کندری*، بهارک ابراهیمی بهنام
    مقدمه

    از جمله دلایل احتمالی برای سقط های مکرر با علت نامشخص، وجود پلی مورفیسم های ژنی می باشد. مطالعه حاضر با هدف بررسی فراوانی و همراهی پلی مورفیسم های rs1233334 و rs2249863 بالادست ژن HLA-G با سقط مکرر خودبه خودی در جمعیت زنان شمال غرب ایران انجام شد.

    روش کار

    این مطالعه مورد- شاهدی در سال های 98-1397 بر روی 160 نفر از زنان مراجعه کننده از منطقه شمال غرب ایران به مرکز ناباروری مادر واقع در شهر تبریز انجام گرفت. 80 نفر از آنها با سابقه سقط مکرر خودبه خودی (حداقل 2 سقط) به عنوان گروه آزمایش و 80 نفر دیگر دارای حداقل یک فرزند و بدون سابقه سقط به عنوان گروه کنترل بودند. پس از تهیه نمونه خون محیطی از آنها، DNA ژنومی استخراج و پلی مورفیسم های مذکور به کمک PCR و تکنیک Sequencing بررسی شدند. تجزیه و تحلیل داده ها با استفاده از نرم افزار آماری SPSS (نسخه 22) و آزمون فیشر انجام شد. میزان p کمتر از 05/0 معنادار در نظر گرفته شد.

    یافته ها

    فراوانی های آلل G و ژنوتیپ GG جایگاه پلی مورفیسم rs2249863 بین دو گروه بیمار و کنترل تفاوت آماری معنی داری نشان داد (05/0<p). در مورد جایگاه پلی مورفیسم rs1233334، فراوانی های آلل C و ژنوتیپ CC هرچند بین دو گروه بیمار و کنترل متفاوت بودند، اما این تفاوت ها از نظر آماری معنی دار نبودند (05/0>p). همچنین بررسی هاپلوتایپی دو جایگاه، شایع ترین هاپلوتایپ را هاپلوتایپ CG مشخص کرد که فراوانی آن بین گروه های بیمار و کنترل از نظر آماری معنی دار بود (05/0<p).

    نتیجه گیری

    پلی مورفیسم rs2249863 و هاپلوتایپ CG جایگاه های rs2249863 و  rs1233334با اختلال سقط مکرر خودبه خودی در جمعیت مورد مطالعه همراهی نشان می دهند و می توان آنها را به عنوان فاکتور خطر بالقوه در نظر گرفت.

    کلید واژگان: پلی مورفیسم rs2249863 و rs1233334, سقط مکرر خودبه خودی, HLA-G
    Leili Khasevani, Mohammad Khalaj Kondori *, Baharak Ebrahimi Behnam
    Introduction

    Genetic polymorphisms are one of the possible causes of recurrent abortion with unknown cause. This study was performed with aim to investigate the association and frequency of rs2249863 and rs1233334 polymorphisms of the upstream region of HLA-G gene with the recurrent spontaneous abortion in women from Northwest of Iran.

    Methods

    This case-control study was performed on 160 women from Northwest of Iran referring to the Madar infertility clinic of Tabriz between 2018 and 2019. Among them, 80 women with a history of recurrent spontaneous abortion (at least 2 abortions) were considered as case group and 80 women with at least one child and without any history of miscarriage as control group. After collecting blood samples, genome DNA was extracted and the polymorphisms were evaluated by PCR and sequencing technique. Data were analyzed using SPSS software (version 22) and Fisher test. P <0.05 was considered statistically significant.    

    Results

    The frequencies of allele G and genotype GG in the rs2249863 polymorphism were significantly different between the case and control groups (P<0.05). In the rs1233334 polymorphism, although the frequencies of allele C and genotype CC were different between the case and control groups, but the differences were not statistically significant (P>0.05). Moreover, haplotype analysis identified the CG as the commonest haplotype that its frequency was significantly different between the case and control groups (P<0.05).

    Conclusion

    The rs2249863 polymorphism and CG haplotype of the rs2249863 and rs1233334 polymorphisms are associated with the recurrent spontaneous abortion in the studied population and they could be considered as potential risk factors for the disease.

    Keywords: HLA-G, Polymorphism, Recurrent spontaneous abortion, rs1233334, rs2249863
  • Esmat Khaleqsefat, Mohammad Khalaj-Kondori *, Mortaza Jabarpour Bonyadi, Hamid Soraya, Behnam Askari
    The requirement of varying doses of warfarin for different individuals can be explained by environmental and genetic factors. We evaluated the frequency of vitamin K epoxide reductase complex subunit 1 (VKORC1) and cytochrome P450 2C9 (CYP2C9) variants together with patientdemographic characteristics and investigated their association with warfarin dose requirement with the objective to suggest a warfarin dosing algorithm. In this study, 185 patients with heart valve replacement from West Azerbaijan, Iran were genotyped for VKORC1 (-1639 G>A) and CYP2C9 (*2 and *3 alleles) by PCR-RFLP. Multiple linear regression was performed to create a new warfarin dosing algorithm. The frequency of variants in studied subjects was 12% for CYP2C9 *2, 25.8% for CYP2C9 *3, and 60% for -1639A. The patients who carried the A allele at position -1639 VKORC1 and the variants CYP2C9 *2 and *3 required a significantly lower daily mean warfarin dosage (P = 0.001). Statistical analysis also indicated a significant relationship between the daily maintenance dose of warfarin with age and blood pressure among the studied patients’ cohort (P < 0.001). This study showed that in the heart valve replacement patients considering VKORC1 and CYP2C9 polymorphisms beside demographic characteristics such as age will be helpful in pre-treatment dosing of warfarin which in turn reduces the complications associated with inappropriate warfarin dosing.
    Keywords: VKORC1, CYP2C9, Genetic Polymorphism, warfarin, Demographic characteristics
  • Asiyeh Jebelli, Mohammad Khalaj-Kondori *, Mohammad Rahmati-Yamchi
    Purpose

    Beta-boswellic acid (βBA) may play central roles in neural plasticity. Neural plasticity has significant implications for learning and memory which are governed by strict memoryrelated molecular pathways. To gain insight into the molecular mechanism by which βBA affects these pathways this study analyzed the expression patterns of Camk2α and Camk4 genes in PC12 cells treated with βBA.

    Methods

    The cytotoxic effects of different βBA concentrations on PC12 cells were examined by MTT assay. For gene expression analysis, cells were treated with concentrations of 1 and 10 µM of βBA for 12, 24, 48, and 72 hours. Total RNA was purified by RNX-Plus solution and reverse transcribed into cDNA using Thermo Scientific Reverse Transcription reagents. The expression patterns of Camk2α and Camk4 genes were quantified by quantitative reverse transcription polymerase chain reaction (qRT-PCR).

    Results

    MTT assay indicated that βBA reduced PC12 cell viability in a time- and concentrationdependent manner. The 50% inhibitory concentrations for the 48 and 72 hours time points were 35 and 26 µM, respectively; while, the βBA concentrations up to 100 µM failed to kill 50% of the cells after 24 hours. According to the qRT-PCR data, the Camk2α variant is not expressed in either βBA-treated or untreated PC12 cells. However, a significant upregulation was observed in Camk4 after 12 hours of treatment with βBA, which followed by a significant downregulation after 24 hours and a persistent expression equal to the control until 72 hours.

    Conclusion

    these findings indicate that PC12 cells not only does not express Camk2α but also its expression cannot be induced by βBA. However, βBA does modulate the expression of Camk4. This result provides further insight into the molecular mechanism by which βBA affects memory.

    Keywords: Beta boswellic acid, Camk2α, Camk4, PC12 cells, Memory
  • یاسمن همپانژاد، محمد خلج کندری*، مجید خدایی، مهناز طالبی
    زمینه و هدف

    بیماری آلزایمر شایع ترین عامل زوال عقلی در افراد مسن است؛ و یک بیماری پیش رونده و برگشت ناپذیر مغزی است. گیرنده سرتیلین در نقل و انتقالات سلولی پروتئین پیش ساز آمیلوئید بتا درگیر است و نیز نقش مهمی در تولید پپتید آمیلوئید بتا در آلزایمر دارد. هدف این پژوهش مطالعه ی همراهی پلی مورفیسم rs2282649 و rs12285364 با بیماری آلزایمر در جمعیت آذری شمال غرب ایران بود.

    روش بررسی

    افراد مورد مطالعه در این پروژه؛ مورد - شاهدی در دو گروه شامل 100 نفر بیمار و 83 نفر سالم به عنوان کنترل وارد شدند جهت تعیین ژنوتیپ ها از تکنیک (PCR-RFLP) استفاده شد.

    یافته ها

    در اسنیپ rs2282649 فراوانی ژنوتیپ CC در گروه بیمار 36٪ و در گروه شاهد 38/55٪ با 0/70= Pبوده و فراوانی ژنوتیپ TT در گروه بیمار 12٪ و در بین گروه شاهد 7/23٪ بود (0/25 =P). فراوانی ژنوتیپ CT در گروه بیمار 52٪ و در گروه شاهد 54/22٪ بود (0/75 =P) و همچنین در rs12285364 فراوانی ژنوتیپ CC در گروه بیمار 93٪ و در گروه شاهد 90/36٪ با 0/5= P بوده و فراوانی ژنوتیپTT  در گروه بیمار و در بین گروه شاهد صفر درصد بود و فراوانی ژنوتیپCT در گروه بیمار 7٪ و در گروه شاهد 9/46٪ (0/49 =P) بود.

    نتیجه گیری

     با توجه به اینکه هر دو گروه در فراوانی ژنوتیپ ها تفاوت معنی داری را نشان ندادند. مشاهده گردید؛ که این پلی مور فیسم ها با بیماری آلزایمر در جمعیت شمال غرب ایران ارتباطی ندارند.

    کلید واژگان: بیماری آلزایمر, پلی مورفیسم, گیرنده سرتیلینSORL1, شمال غرب ایران
    Yasaman Hampanezhad, Mohammad Khalaj Kondori*, Majid Khodayi, Mahnaz Talebi
    Background and Aim

    Alzheimer’s disease is the most common cause of dementia in old people. AD is a progressive and irreversible neurodegenerative brain disorder. Sortilin receptor 1 (SORL1) is involved in cellular trafficking of amyloid precursor protein and plays an essential role in amyloid-beta peptide generation in the AD. The purpose of the present study was to evaluate the association of SORL1 rs2282649 and rs12285364 polymorphism with AD in Azeri population in the northwest of Iran.

    Materials and Methods

    This case- control study included 100 Alzheimer’s disease patients as our case and 83 healthy subjects as our control group. Genotypes were determined by (PCR- RFLP) technique.

    Results

    In rs2282649 SNP the frequency of homozygous CC genotype was 36% in the case and 38.55% % in the control groups (P= 0.70). The frequencies of TT genotype were 12% and 7.23% in the case and control groups respectively (P = 0.25). The frequency of heterozygote CT genotype was 52% in the cases and 54.22% in the control group (P = 0.75). Also, in rs12285364 polymorphism the frequencies of homozygous CC genotype in the cases and the control groups were 93% and 90.36 % respectively (P= 0.5). The frequency of TT genotype was 0% in both case and control groups (P = 0.00). The frequencies of heterozygote CT genotype were 7% in the case and 9.64% in the control groups (P = 0.49).            

    Conclusion

    No statistically significant difference was observed between the case and control groups in regard to the frequencies of the genotypes. Therefore we can conclude that these polymorphisms are not associated with Alzheimer’s disease among the people in the northwest of Iran.

    Keywords: Alzheimer’s disease_polymorphism_Sortilin receptor 1 (SORL1)_Northwest of Iran
  • Elham Mehdizadeh, Mohammad Khalaj Kondori, Zeinab Shaghaghi Tarakdari, Saeed Sadigh Eteghad, Mahnaz Talebi*, Sasan Andalib
    Introduction

    Alzheimer’s disease (AD), which is a progressive neurodegenerative disorder, causes structural and functional brain disruption. MS4A6A, TREM2, and CD33 gene polymorphisms loci have been found to be associated with the pathobiology of late-onset AD (LOAD). In the present study, we tested the hypothesis of association of LOAD with rs983392, rs75932628, and rs3865444 polymorphisms in MS4A6A, TREM2, CD33 genes, respectively.

    Methods

    In the present study, 113 LOAD patients and 100 healthy unrelated age- and gendermatched controls were selected. DNA was extracted from blood samples by the salting-out method and the genotyping was performed by RFLP-PCR. Electrophoresis was carried out on agarose gel. Sequencing was thereafter utilized for the confirmation of the results.

    Results

    Only CD33 rs3865444 polymorphism revealed a significant difference in the genotypic frequencies of GG (P=0.001) and GT (P=0.001), and allelic frequencies of G (P=0.033) and T (P=0.03) between LOAD patients and controls.

    Conclusion

    The evidence from the present study suggests that T allele of CD33 rs3865444 polymorphism is associated with LOAD in the studied Iranian population.

    Keywords: Late onset Alzheimer’sdisease, LOAD, MS4A6A, CD33, TREM2, Polymorphisms
  • Asiyeh Jebelli, Mohammad Khalaj Kondori *, Mortaza Bonyadi, Mohammad Ali Hosseinpour Feizi, Mohammad Rahmati Yamchi
    Physiological studies confirm improvement of memory by Olibanum, a resin from Boswellia species, while little is known about the molecular mechanism by which it affects memory performance. Two master transcription factors, CREB-1 and CREB-2, regulate downstream memory-related genes expression, leading to the long-term memory potentiation. This study addresses the effects of Beta-boswellic acid (β-BA), the main ingredient of Olibanum, and ethanolic extract of the resin from Boswellia serrata on the expression of CREB-1 and CREB-2 genes in B65 cell line. B65 cells were treated with β-BA) or ethanolic extract of Olibanum in different dose and time intervals and the cell viability/toxicity was measured by MTT assay. Total RNA was extracted from the treated and untreated control cells and cDNA was synthesized. The expression levels of CREB-1 and CREB-2 genes were quantified by Real-time PCR. MTT assays revealed 50% inhibitory concentrations of 42.05, 29.63 and 21.78 μg/ml for ethanolic extract of Olibanum and 89.54, 44.05 and 21.12 µM for β-BA at 24, 48 and 72h time intervals respectively. Both β-BA and ethanolic extract of Olibanum altered CREB-1 and CREB-2 genes expression levels in time-dependent but not in dose-dependent way. However, β-BA showed stronger and more stable effects. The expression levels of the both genes followed an alternate upregulation and downregulation pattern, but in opposite directions, in response to the both solutions with the progress of time. These results suggest that Olibanum possibly improves memory performance, at least partially, by regulating the levels of CREB-1 and CREB-2 transcription factors via positive/negative-feedback loops.
    Keywords: memory, Olibanum, Beta-boswellic acid, CREB-1, CREB-2
  • آسیه جبلی، محمد خلج کندری *، مرتضی بنیادی، محمدعلی حسینپور فیضی، محمد رحمتی یامچی
    هدف
    مهم ترین تاثیرات کندر در ارتباط با حافظه عبارت است از پیشگیری یا بهبود بیماری آلزایمر و نیز تقویت حافظه زاده های رت هایی که در دوران بارداری و شیردهی کندر دریافت کرده اند. ژن Camk4 یکی از ژن های مهم حافظه می باشد که با فسفریلاسیون و فعال کردن فاکتورهای رونویسی بیان ژن های پایین دست حافظه را القا می کند. بر این اساس، هدف این مطالعه بررسی تاثیر عصاره آبی کندر در بیان ژن Camk4 در دو رده سلولی PC12 و B65 می باشد.
    مواد و روش ها
    تاثیر عصاره بر زیستایی سلول ها با تیمار سلول ها در دو بازه زمانی و 6 غلظت مختلف عصاره بررسی شد و غلظت مهاری 50 درصد به دست آمد. برای مطالعات بیان ژن، سلول ها با دو غلظت عصاره در دو بازه زمانی تیمار شدند. سپس RNA سلولی استخراج شده و به cDNA تبدیل شد. در نهایت، واکنش qPCR جهت بررسی بیان ژن Camk4 در سلول ها انجام شد.
    یافته ها
    عصاره آبی کندر زیستایی سلول ها را در یک الگوی وابسته به زمان و غلظت کاهش داد. با این وجود، سمیت عصاره برای رده سلولی B65 بیش تر از PC12 بود. هم چنین، عصاره کندر توانست بیان ژن Camk4 را به طور قابل توجهی در سلول ها افزایش دهد. با این وجود، این افزایش وابسته به غلظت و نوع سلول بود و بیان ژن Camk4 در غلظت کم عصاره و در رده سلولی PC12 بیش تر و سریع تر از B65 افزایش یافت.
    نتیجه گیری
    نتایج این مطالعه نشان می دهد کندر در یک الگوی وابسته به غلظت می تواند بیان ژن Camk4 را تنظیم کند. با این وجود، به مطالعات تکمیلی بیش تری جهت شناسایی مکانیسم های عمل کندر در سلول های مختلف نیاز است.
    کلید واژگان: کندر, حافظه, ژن Camk4, رده سلولی PC12
    Asiyeh Jebelli, Mohammad Khalaj-Kondori *, Mortaza Bonyadi, Mohammad Ali Hosseinpour Feizi, Mohammad Rahmati-Yamchi
    Introduction
    The most important effects of Frankincense on memory are included prevention and relative treatment of Alzheimer disease, as well as memory enhancement of off springs of the rats that had received Frankincense during their pregnancy and lactation period. Considerably, Camk4, one of the important memory genes, induces downstream memory genes expression through phosphorylation and activation of transcription factors. Here, we aimed to study the effects of aqueous extract of Frankincense on the Camk4 gene expression in PC12 and B65 cell lines.
    Materials And Methods
    The effect of extract on the cell viability was evaluated by cell treatment in two time intervals with six concentrations of extract and 50% inhibition concentration was obtained. In this way, for gene expression studies, cells were treated by two concentrations of extract in two time points. RNA was extracted and converted to cDNA and qPCR was performed to investigate the expression of Camk4 gene.
    Results
    The aqueous extract of Frankincense decreased the cell viability in a time and concentration dependent pattern. However, the extract was more toxic for B65 cell line than PC12. Also, it significantly increased the expression of Camk4 gene in the cells. Nevertheless, the increase was dependent to the extract concentration and the cell type. The low concentration of the extract was more effective to the expression of Camk4 gene than the high concentration as well as to its expression in the PC12 cell line than B65.
    Conclusion
    This study showed that Frankincense could regulate the expression of Camk4 gene in a concentration dependent pattern. However, further studies are needed to identify the mechanisms of Frankincense action in different cells.
    Keywords: Frankincense, Memory, PC12 cell line, Camk4 gene
  • کبری احمدی ثانی، محمد خلج کندری*، محمدعلی حسینپور فیضی
    سابقه و هدف
    کندر از جمله مواد گیاهی می باشد که خواص درمانی مختلفی برای آن ذکر شده است. بتا-بوسولیک اسید، ترکیب اصلی کندر، حلالیت کمی در آب دارد و دسترسی زیستی آن بسیار پایین می باشد. هدف این مطالعه بررسی اثر بارگذاری بتا- بوسولیک اسید در نانوذره های دندروزومی بر میزان دسترسی زیستی و جذب سلولی آن می باشد.
    مواد و روش ها
    برای انجام آزمون MTT، سلول های B65 با غلظت های مختلف بتا- بوسولیک اسید تنها و بتا- بوسولیک اسید دندروزومی به مدت 24، 48 و 72 ساعت تیمار شدند. پس از افزودن MTT، شدت رنگ تولید شده توسط ELISA reader اندازه گیری و IC50 محاسبه شد. داده های حاصل با استفاده از برنامه آماری One Way Anova و نرم افزار SPSS v.16 آنالیز شدند.
    یافته ها
    مقدار IC50 برای بتا- بوسولیک اسید تنها و بتا- بوسولیک اسید دندروزومی به ترتیب 09/88 و 42/58 میکرومولار برای 24 ساعت، 37/58 و 87/44 میکرومولار برای 48 ساعت و 09/21 و 69/16 میکرومولار برای 72 ساعت به دست آمد.
    نتیجه گیری
    نتایج حاصل نشان می دهد که بارگذاری بتا-بوسولیک اسید در نانوذره های دندروزومی باعث افزایش سمیت سلولی بتا-بوسولیک اسید می شود که این مشاهده می تواند به علت افزایش جذب سلولی بتا-بوسولیک اسید در حضور نانوذره های دندروزومی باشد.
    کلید واژگان: بتا- بوسولیک اسید, دندروزوم, تست MTT, رده سلولی B65
    Kobra Ahmadi Sani, Mohammad Khalaj Kondori *, Mohammad Ali Hosseinpour Faizi
    Aim &
    Background
    Frankincense is a herbal product that has diverse therapeutic effects. Beta-boswellic acid, the main ingredient of frankincense, is poorly soluble in water and its bioavailability is very low. The aim of present study was to evaluate the effect of loading of beta-boswellic acid in dendrosomal nanoparticles on its bioavailability and cell uptake by MTT assay.
    Materials And Methods
    For MTT assay, B65 cells were treated with different doses of nave beta-boswellic acid or dendrosomal beta-boswelic acid for 24, 48 and 72 hours. After adding MTT, colorimetery was done using ELISA reader and IC50 was then calculated. The data was analyzed with One Way Anova program using SPSS v.16 software.
    Results
    The IC50 for nave beta-boswellic acid and dendrosomal beta-boswellic acid was 88/09 and 58/42 µM in 24h, 58/37 and 44/87 µM in 48h and 21/09 and 16/69 µM in 72h respectively.
    Conclusion
    The results showed that loading of beta-boswellic acid in dendrosomal nanoparticles increases the cell toxicity effect of beta-boswellic acid. This observation might be due to the increased cell uptake of beta-boswellic acid in the presence of dendrosomal nanoparticles.
    Keywords: beta- Boswellic acid, Dendrosome, MTT test, B65 cell line
  • محبوبه کاووسی، محمد خلج کندری*، مهدی کدیور
    زمینه و اهداف

    انتقال هدفمند ناقل ها و حامل های ژنی به سلول ها و بافت های خاص یکی از مهمترین مسائل در آزمایش های ژن درمانی است. باکتریوفاژها حامل-های ژنی مناسبی هستند و روش های مختلفی برای انتقال هدفمند آن ها به سلول های مورد نظر وجود دارد. در این مطالعه تهیه ذرات فاژی هدفمند شده بر پایه فاژ M13 به روش اتصال شیمیایی ارائه می شود.

    مواد و روش ها

    ابتدا سازه فاژمیدی pCMV-Script-GFP از ذرات فاژی GFP توسط فاژ کمکی ExAssist به روش برش درون سلولی با استفاده از سویه میزبان اختصاصی XLOLR برش و با استفاده از فاژ کمکی R408 ذرات فاژیM13حاوی ژن GFP (M13-GFP)تولید شدند. سپس ملکول های پروتئین هولوترانسفرین انسانی با روش اتصال شیمیایی آمیناسیون احیایی به سطح ذرات فاژی حاصل اتصال داده شدند. اتصال ملکول های ترانسفرین به سطح ذرات فاژی با روش phage-ELISA بررسی شد.

    یافته ها

    آزمایشات phage-ELISA نشان داد که اتصال ملکولهای ترانسفرین به سطح ذرات فاژی به درستی انجام گرفته و ذرات فاژیM13 هدفگیری شده با ترانسفرین تهیه شده است. بررسی های بیشتر نشان داد که تقریبا 485 مولکول ترانسفرین به هر ذره فاژی متصل شده است.

    نتیجه گیری

    نتایج حاصل نشان می دهند که روش اتصالات شیمیایی می تواند به عنوان راهکاری مناسب در هدفمندسازی ذرات فاژی M13 با اتصال مولکول های هدفگیری به تعداد زیاد به سطح آن ها مورد استفاده قرار گیرد.

    کلید واژگان: اتصال شیمیایی, فاژ M13, هولوترانسفرین, هدفگیری
    Mahboubeh Kavooci, Mohammad Khalaj Kondori*, Mehdi Kadivar
    Background and Objectives

    Targeting transgene carriers and vectors to individual cells and tissues is one of the most important goals of gene therapy. Bacteriophages are of appropriate transgene carriers and there are different methods for their targeting to target cells. Present study reports preparation of targeted M13-based bacteriophage particles by a chemical coupling strategy.

    Materials And Methods

    First, the pCMV-Script-GFP construct was produced via in vivo excision protocol from GFP Phage particles using ExAssist helper phage and XLOLR as specific host. Then, M13 phage particles bearing GFP (M13-GFP) were obtained by single stranded rescue using R408 helper phage. The human holotransferrin molecules were then coupled to the surface of phage particles by reductive amination chemistry. Transferrin molecules bind to the surface of phage particles were studied by phage-ELISA.

    Results

    Phage-ELISA tests showed that holotransferrin molecules were coupled to the surface of M13 phage particles in a correct way and the transferrin-targgeted M13 phage particles were prepared. Further analysis showed that about 485 transferrin molecules coupled per phage particle.

    Conclusion

    The results show that chemical coupling might be considered as a suitable strategy for targeting of M13 particles via coupling of targeting molecules in high density to the phage surface.

    Keywords: Chemical Coupling, M13 Phage, Holotransferrin, Targeting
  • Elham Abedheydari, Mohammad Khalaj-Kondori, Mohammad-Ali Hosseinpour-Faizi, Morteza Kosari-Nasab
    Gene delivery might be affected by several tribulations based on carrier/vector applied. Bacteriophages lambda and M13 have different genome conformations; linear double-stranded and circular single-stranded respectively. Therefore, it might be expected that these two common classes of gene delivery vehicles will have different capacity for gene delivery and expression in eukaryote cells. To address the possible effects of linear double-stranded and circular single-stranded genome conformations of bacteriophages lambda and M13 on the transgene expression, the transfection efficacy of two vectors based on lambda and M13 were compared in AGS cell line. The GFP encoding sequence was inserted into the Lambda ZAP-CMV XR vector which resulted in λ-ZAP-CMV-GFP construct. The construct was then in vitro packaged using Gigapack® III Gold packaging extract and λ-GFP phage particles were obtained. The λ-GFP phage particles were then used for in vivo excisioning which resulted in M13-CMV-Script-GFP construct. 1011 copy of λ-ZAP-CMV-GFP or M13-CMV-Script-GFP constructs were transfected into AGS cells using lipofectamine 2000. Transfection efficiencies were analyzed by FACS. Results showed that linear double-stranded λ-ZAP-CMV-GFP was efficient than single-stranded form of M13-CMV-Script-GFP while its double-stranded form was efficient than the linear double-stranded λ-ZAP-CMV-GFP construct for transgene delivery and expression. Moreover the GFP signals resulted from transfections by single-stranded form of M13-CMV-Script-GFP construct faded more quickly in comparison to others. These findings highlight that genome conformation of gene carriers might be an important factor when seeking for an appropriate gene carrier/vehicle.
    Keywords: gene delivery, phage, mediated gene transfection, vector conformation
  • محمد علی حسینپور فیضی، مرضیه قنبریان، محمد خلج کندری، ثریا شیری، وحید منتظری، منیره حلیمی
    زمینه و هدف
    تنظیم بیان ژن در سطح ترجمه به ویژه در مرحله آغازین وابسته به عملکرد فاکتور آغازگر یوکاریوتی «eIF4E»در قالب کمپلکس آغازگر EIF4F می باشد. گزارشات مبنی بر بیان متفاوت و بالای ژن EIF4E در چندین سرطان از جمله سینه، روده، ریه، سر و گردن، پروستات و سیستم هماتوپوئتیک، مؤید نقش این فاکتور در تومورزایی و پیشرفت سرطان است. لذا در این تحقیق تلاش بر این بوده است که میزان بیان این فاکتور پروتئینی به عنوان یک مارکرمولکولی جدید در تومورهای تیروئید و بافت های حاشیه ی آن مورد ارزیابی قرارگیرد.
    روش بررسی
    در تحقیق حاضر 21 نمونه توموری از نوع پاپیلاری و 14 نمونه حاشیه ی تومور به صورت یک مطالعه ی مورد شاهدی با استفاده از تکنیک Reverse transcriptase polymerase chain reaction (RT-PCR)به صورت نیمه کمی مورد بررسی قرار گرفت و ژن ß2m جهت نرمال سازی نتایج به کار گرفته شد. به منظور آنالیز آماری، میزان بیان eIF4Eبا استفاده از نرم افزار SPSS (نگارش 16) و تست های آماری پارامتریک One way-Anova و t-test در گروه های مختلف مورد مقایسه قرار گرفته شد.
    یافته ها
    بررسی ها نشان داده است که 1) ژن eIF4E در تمامی نمونه های مورد مطالعه بیان شده است.2) میزان بیان این فاکتور در نمونه های سرطانی نسبت به نمونه های حاشیه ی توموری افزایش یافته است. 3) سطوح بیان این ژن در بافت های بدخیم سرطانی بیشتر از بافت های خوش خیم بوده است.
    نتیجه گیری
    در مجموع یافته های این تحقیق نشان داد که ژن eIF4E به عنوان فاکتور آغازگر ترجمه ای دارای بیان سراسری بوده و میزان بیان آن با ماهیت سرطانی تومور ها و درجه ی بدخیمی تومو رهای سرطانی ارتباط مستقیمی داشته است. این نتایج ضمن تایید نقش ژن eIF4E در پیشبرد روند سرطانی، افزایش بیان این ژن را در بروز تومورهای تیروئیدی مؤثر دانست. بنابراین ارزیابی بیان ژن eIF4E می تواند ملاک مناسبی جهت تمایز بیماری های توموری از انواع غیر توموری محسوب شود و به عنوان مارکری جهت رده بندی تومورهای تیروئیدی مورد استفاده قرار گیرد. این فاکتور پیش آگهی و تشخیصی، همچنین می تواند هدف درمانی نوید بخش برای درمان سرطان باشد.
    کلید واژگان: سرطان تیروئید, eIF4E, پیشرفت سرطان, RT, PCR نیمه کمی
    Dr Mohammad Ali Hoseinpourfeizi, Marzieh Ghanbarian, Mohammad Khalaj-Kondori, Soraya Shiri, Dr Vahid Montazeri, Dr Monireh Halimi
    Background And Aim
    Regulation of protein synthesis in the early stage of translation depends on the function of eIF4E factor especially in the form of eukaryotic initiation factor (eIF4F). Overexpression of eIF4E in multiple cancer types, including malignancies of the prostate, breast, colon, lung, and the hematopoietic system is indicative of the role of this factor in tumorogenesis and promotion of the cancers. In this study we investigated the expression pattern of eIF4E as a new molecular marker in thyroid tumors and their marginal normal tissues.
    Materials And Methods
    We used semi-quantitative RT-PCR technique to examine the expression of eIF4E in 21 papillary carcinoma tissue specimens and 14 specimens of corresponding marginal normal tissue adjacent to the malignant lesions. ß2m gene was considered as an internal control. Rate of expression of eIF4E in different groups were compared with one another by use of SPSS software and data were analyzed by one way ANOVA and t-test.
    Results
    Our data revealed significant expression of eIF4E in all tumor samples compared to non-tumor lesions and normal tissues (P<0.05). Moreover the expression level was notably increased in malignant tumor samples compared to marginal tissues of the tumors (P<0.05). The rate of expression was more in tumor samples than non-malignant samples.
    Conclusion
    The results of this study indicated that the rate of expression of eIF4E gene is associated with kind of tumor and grade of malignancy. Also this study confirmed the role of eIF4E gene in tumor progression and development of thyroid tumors. Therefore eIF4E gene expression can be an appropriate indicator for diagnosis of tumors and can be used as a guide for grading of thyroid tumors. This prognostic and diagnostic factor can be considered as a promising therapeutic target for treatment of cancers.
سامانه نویسندگان
  • دکتر محمد خلج کندری
    دکتر محمد خلج کندری

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