جستجوی مقالات مرتبط با کلیدواژه "qrt-pcr" در نشریات گروه "پزشکی"

The We evaluate the effects of radiofrequency electromagnetic field (RF-EMF) on rat brain and testicular tissue using histopathology, comet assay, and real-time quantitative PCR techniques. Two equal groups of fourteen rats one for sham-control and the other for exposure (n = seven) were created. For a duration of 14 days, the exposure group (2100 MHz, testicular tissue SAR values of 163 mW/kg for 10 g, brain tissue SAR values of 292 mW/kg on average) was subjected to five hours of exposure per day. Evaluations were conducted on tissue gene expression levels, histopathology, and DNA damage to brain tissue. The histological examination of brain tissue from the exposed group revealed vascular alterations and significant edema (p < 0.05). It was determined that RF radiation-induced much more cellular damage in the exposed group (18.26% tail DNA) than in the control group (4.06% tail DNA). Signs of deterioration in spermatogenic cells in the testicular tissue of the exposed group also changed significantly (p < 0.05). The Bax and bcl-2 genes showed a significant difference (p < 0.05) in the mRNA level data, whereas the p53 genes showed no significant change (p > 0.05). These findings suggest that it may cause some histopathological and cellular damage in brain and testis tissue.

A critical role has been known for lncRNAs in the initiation and development of cancers. Therefore, lncRNAs have been reported as the possible biomarkers in relation to the diagnosis and therapy of malignancies. This project examined the change in CYTOR lncRNA expression in human cervical cancer samples as compared with adjacent healthy ones.

We provided one hundred fifteen pairs of tumorous and adjacent healthy tissue specimens of cervical cancer patients. RNAs were isolated from tissue specimens and cDNAs were synthesized. We considered quantitative Real-time PCR (qRT-PCR) to examine the expression levels of CYTOR lncRNA. In addition, the biomarker activity of CYTOR and the associations between the lncRNA and clinicopathological characteristics were evaluated.

The significant increased expression of CYTOR was obtained in cancerous samples as compared with non-cancerous ones (P< 0.0001). A significant correlation was indicated between CYTOR expression and the squamous subtype of cervical cancer (p=0.046). The receiver operating characteristic (ROC) curve-related AUC (area under the curve), specificity, and sensitivity were calculated 0.88, 81.74%, and 80%, respectively, which may introduce CYTOR as a potential biomarker.

CYTOR may be an effective oncogene and biomarker in cervical cancer cases given its increased expression in human cervical cancer tissues.

In recent years, viral epidemics such as the coronavirus disease (COVID-19) have spread, and this outbreak is thought to be the result of animal-to-human transmission. Hence, accurate diagnostic tests to detect COVID-19 and antiviral antibodies in infected individuals are of utmost importance. This report describes the structure, history, taxonomy, and molecular and immunological techniques for diagnosing this disease. Tests for early diagnosis of COVID-19 depend on the reverse transcription-polymerase chain reaction (RT-PCR). However, tests based on isothermal amplification and clustered regularly interspaced short palindromic repeats (CRISPR)-based methods are promising options. Identifying people whose activated antibodies require serological tests, including enzyme-linked immunosorbent assays (ELISA). The search for efficient, cost-effective, and accurate laboratory techniques that can be used on a large scale continues. The RT-PCR technique is a dominant technique for the detection of viral RNA. Other acidic nucleic-based assays such as isothermal amplification, microarray hybridization, amplicon-based metagenomics sequencing, and CRISPER-based techniques have been developed. Along with molecular methods, different efficient serological and immunological methods such as ELISA, rapid antigen test, lateral flow immunoassay, luminescent Immunoassay, and biosensors are also developed.

The occurrence of gastric cancer is associated with numerous aspects, including the host's lifestyle and genetic history. Understanding gastric cancer molecular mechanisms can improve our insight into the early diagnosis, prognosis, and treatment. In this study, the RNA level of SNHG8, AF147447, and n34560 genes in gastric tumor tissues was investigated and their association with Helicobacter pylori and Epstein-Barr virus infections was evaluated. Formalin-fixed paraffinembedded (FFPE) tissues (100 samples), including 50 samples of gastric cancer tissues and 50 samples of healthy tissues were taken. The expression level of SNHG8, AF147447, and n34560 genes in gastric cancer and control tissues were examined using the qRT-PCR technique. A significant association was observed between the expression level of the SNHG8 gene in gastric tumor tissues compared to the healthy tissues (P=0.0003). Relative expression of AF147447 and n34560 genes did not show any significant difference among gastric tumor tissues compared to the normal tissues (P=0.2984, P=0.9158). In addition, pathological comparison of clinical data with the expression of SNHG8, AF147447, and n34560 genes did not show any significant association in tumor and healthy tissues, but the expression level of AF147447 gene in Helicobacter pylori infection (P=0.0458) and expression level of n34560 gene in Epstein-Barr virus (EBV) infection (P=0.0362) showed significant association. In conclusion, we found a significant association between SNHG8 gene expression levels and the possible cancer incidence. Also, a significant association was observed between the expression of n34560 and AF147447 genes relating to H.pylori and EBV infections in gastric cancer.

GD2 synthase (GD2S) is the key enzyme required for ganglioside GD2 synthesis. It is commonly expressed in normal tissues and various cancers. Ganglioside GD2 is identified as a breast cancer stem cells (BCSCs) marker that promotes tumorigenesis. As GD2S has been found to be a useful molecular marker in neuroblastoma and retinoblastoma tumors, we suggest that it can be considered as a suitable candidate for the detection of CSCs in breast cancer tissues.

Expression of GD2S was examined in 65 breast tumors compared to adjacent normal tissues, applying quantitative real-time PCR (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). The association between GD2S expression level and patients’ clinical characteristics was also assessed.

Our findings showed that GD2S mRNA expression was significantly higher in breast cancer tissues in comparison to normal adjacent tissue samples (4.92-fold change, p<0.001) in advanced grades (p<0.001) and stages (p<0.001). It was also shown that GD2S protein expression was significantly higher in breast cancer tissues in comparison to normal adjacent tissues (4.86-fold change, p=0.010) in advanced grades (p=0.010), stages (p=0.005) and larger tumor size (p=0.002).

The current study showed that increased expression of GD2S in advanced breast cancer potentiates it as a promising tumor marker in these patients.

Spot blotch, caused by Cochliobolus sativus, is most effectively managed using fungicide applications, including triadimefon (TDM) a triazole compound. C. sativus posses a great concern as it might develop resistance against fungicides like TDM due to its high genetic variability, short life cycle, and abundant inoculum production. Therefore, to better understand the mechanisms of TDM resistance initiated by C. sativus, changes in cytochrome b (cytb) gene in virulent and avirulent pathotypes were evaluated at early time points of TDM treatments.

C. sativus sensitivity to TDM was determined by measuring the radial growth of each pathotype on PDA plates. Additionally, RNA was isolated from mycelia of each pathotype at 24, 48, 72 and 96 hours post fungicide treatments and used for cDNA synthesis. Cytb was verified using quantitative reverse transcriptase PCR (qRT-PCR).

Data showed that the maximum mycelial growth inhibition by 50% (EC50) for both pathotypes was recorded 48h at 0.25 µg/ml TDM treatment. The qRT-PCR revealed that cytb expression increased in both virulent and avirulent pathotypes at 24h post TDM treatments in comparison with non-treated controls. The most outstanding differences in cytb expression were7.69 and 2.88-fold in the virulent and avirulent pathotypes, respectively, 48h of 0.25 µg/ml TDM treatment.

According to findings, it is possible to propose that cytb gene might play a role in signaling events during C. sativus exposure to commercial triazole fungicide.

Epithelial ovarian cancer (EOC), is the fatal form of gynecological cancer. Almost 70% of ovarian cancer patients are detected at an advanced stage (III-IV) with metastases. Epithelial‑mesenchymal transition (EMT) is a critical process associated with metastasis. This study investigated the expression levels of AXL, GAS6, Claudin-1, and Cofilin-1, as genes involved in EMT in relation to clinicopathologic features in ovarian cancer patients.

In this descriptive study, 78 ovarian epithelial cancer patients were enrolled. Samples were provided by the Iran National Tumor Bank, founded by the Cancer Institute of Tehran University of Medical Sciences in 2017. The expression levels of AXL, GAS6, Claudin-1, and Cofilin-1 genes were investigated in a fresh, frozen tumor sample and normal adjacent tissue by real-time PCR (RT-PCR).

Findings showed a significant relationship between the overexpression of AXL and TNM staging (P=0.03). The expression level of GAS6 decreased in more advanced stages (P=0.01). There is a negative relationship between Cofilin-1 expression level and TNM staging (P=0.002). Claudin-1 expression level was higher in low stages compared with that in high stages (P=0.01). There was no relationship between gene expression levels of target genes with size and grade of the tumor.

Given the importance of these genes in EMT, alteration in their expression pattern can contribute to the progression of the disease and distant metastasis of cancer cells. Additionally, knowing the alteration pattern of these genes expression can help to better understanding and prediction of the prognosis of EOC.

Oral squamous cell carcinoma (OSCC) is the most abundant dysplasia in the oral cavity that aberrant expression of microRNA plays an important role in cancer progression. It is well known that microRNA-21 expression is oncogenes or tumor suppressor factor in malignancy formation of several cancers. In this study used OSCC tissues to investigate the expression of mir-21 in malignancy causes.

OSCC tissues and normal samples were collected from Amir Alam and Taleghani Hospitals as fresh and frozen samples. The Expression level of miR-21 in OSCC and normal tissues were measured by qRT-PCR. To find the targets related mir-21 used databases including TargetScan, GenTrail2, GO and STITCH online websites.

MiR-21 found significantly up-regulated in OSCC tissues compared to normal tissues (Fold-change=5.54). Targets of miR-21 derived from the TargetScan and GeneTrail2 analysis determined the most significant biological processes be associated with the epithelialization, differentiation, and morphogenesis. So overexpression of miR-21 could reverse this process and promote the cells to stemness and metastatic state. EGFR and PDCD4, two targets of miR-21, previously have been demonstrated that are important in OSCC invasion, metastasis, and differentiation. In addition study on targets genes to find anticancer drugs enriched CI-1033 as a tyrosine-kinase inhibitor that previously reported for the treatment of cancer.

Our findings indicate miR-21 act as oncomiR in OSCC and may be considered as a biomarker for the development of OSCC treatment.

Turnip mosaic virus (TuMV) has a wide host range and no resistant commercial canola variety to this virus has been reported in Iran. Thus, RNA silencing mechanism was applied to consider the possibility of improvement in resistance to TuMV in spring canola, RGS003 variety.

To obtain an effective construct for silencing, based on the bioinformatics analysis, a fragment containing 130 conserved nucleotide sequences of the TuMV coat protein gene was gained as targeting candidate to produce sense, antisense and hairpin constructs and assessed for resistance efficiency in a transient expression system in Nicotiana benthamiana by agroinfiltration. The development of symptoms after virus inoculation revealed that the highest efficiency can be obtained by hairpin construct. Therefore, the hairpin construct was applied for the transformation of canola RGS003 via cotyledonary explants using Agrobacterium tumefaciens LBA4404. In transgenic and non-transgenic canola plants, the infection and virus titer were assessed by ratio of detection via ELISA and real-time PCR. In addition, severity of disease symptoms was scored four weeks after inoculation with a TuMV isolate.

Results indicated 5-12 days delay in appearance of symptoms in transgenic plants and there was a decrease in severity of symptoms in contrast to non-transgenic plants. The increased virus concentration ratio in non-transgenic compared to transgenic plants was confirmed by qRT-PCR. The ELISA results confirmed absence of infection on five out of six transformed plants 15 dpi.

These preliminary results proved that transgenic canola plants containing hairpin of 130 nucleotide sequences of TuMV CP gene could resist against TuMV.