جستجوی مقالات مرتبط با کلیدواژه "real- time pcr" در نشریات گروه "علوم پایه"

Gastric cancer is one of the most common cancers in the world. Its treatments are costly and can cause severe side effects. As a result, treatments with natural compounds, well-established therapeutics, or combinations of both groups may be effective alternatives. p-Coumaric acid (pCA) and metformin (Met) are among such anticancer treatments. Epithelial-mesenchymal transition (EMT) is a multi-purpose process that plays a critical role in gastric cancer. This process involves a complex network of biological markers participating in gastric cancer initiation and metastasis. Subsequently, the agents downregulating the expression of EMT markers may be potential anti-gastric cancer therapeutics. Because the effects of pCA, Met, and their combination on the expression of EMT markers ZEB1, Snail2, Vimentin, and VEGFA have not been inspected, the present study aimed at assessing these effects. MTT assay determined the cytotoxicity of pCA and Met on the AGS cells for 48 hours. Real-time PCR was used to evaluate the changes in the expression levels of these EMT genes after 48 hours. A combination of pCA and Met downregulated the expression of ZEB1 and Vimentin genes at low, non-cytotoxic concentrations. Therefore, they may be potential candidates for further investigations in fighting against gastric cancer.

The level of an immune response inhibitory enzyme, called indoleamine 2,3-dioxygenase (IDO), and the activity of Treg cells increase in some patients with cancer and acute leukemias such that it can lead to the inhibition of immune responses. Therefore, the very aim of the present study was to evaluate the regulatory T cells (CD4 + / FOXP3 +) and the level of the IDO expression in acute leukemias using flow cytometry and Real-Time PCR.  

This study used bone marrow samples taken from  30 patients with acute lymphoid leukemia (ALL), and 20 healthy individuals. Then, the level of the IDO expression and the percentage of Treg cells were evaluated using flow cytometry and Real-Time PCR. The results revealed that the percentage of Treg in the total lymphocytes, T and Th in the ALL group was significantly higher than that of the normal groups the respectively (P <0.001).

In addition, the level of the IDO expression in the normal group was significantly lower than that of the ALL group (P= 0.016).

The level of the IDO expression in the ALL groups was significantly higher than the normal group. Moreover, there was a positive and non-significant relationship between the level of the IDO expression and the percentages of Treg cells in the ALL group. Therefore, it seems necessary to conduct more studies for the behavior of this leukemia in the expression of IDO enzyme and the percentage of Treg cells and their relationship with each other.

Brucellosis is one of the most common bacterial diseases in both humans and animals.It causes human to suffer from Malta fever and has highly economic losses in livestock. Bacteriological culture and serological tests for the diagnosis of chronic cases are insensitive. Therefore, the use of Real Time PCR is a great help to evaluate the frequency of types of brucella abortus and brucella melitensis in serum of people with positive serology. Serum samples of 30 cases with right positive test (over 1/160) were collected from DNA clinical laboratories in the cities of Shahrekord and Esfahan and they were extracted with the use of signagen DNA extraction kit. In order to identify the genus and the types of brucella abortus and brucella melitensis Real Time PCR reaction was carried out on the samples. In this study, all 30 samples were confirmed with the use of Real Time PCR. Brucella abortus was detected in all samples in the cities of Esfahan and Shahrekord. After Real Time PCR test was run on females, brucella abortus was just detected, While in males, 22 (%73/33) cases suffered from brucella abortus and 3 (%10) suffered from brucella melitensis. From the total of 7 cases with brucellosis infection in the age group below 30 years old, 6 (%85/7) cases suffered from brucella abortus and 1 (%14/3) suffered from brucella melitensis, While from the 23 cases with brucellosis in the age group over 30 years old, 21 (% 91/3) had brucella abortus and 2 (%8/7) had brucella melitensis. In recent years many molecular methods have been used for the detection of this bacteria. Real Time PCR method in the sense that it requires less time and has high sensitivity and specificity is very useful in the diagnosis of the disease

Brucella spp are small, immobile, gram-negative bacteria that are lacking capsules and formed like cocobacillus. Brucellosis is a zoonosis infection between humans and animals that can lead to miscarriages, fertility disorders, genital infections, reduction in milk production, urethritis, and epididymitis in the original host, resulting in many medical disorders in patients and unnecessary treatment expenses. Moreover, farmers would end up facing significant economic losses. Despite advances in blood culture techniques and serological tests to detect specific antibiotics, there are still significant difficulties in the diagnosis of brucellosis; therefore, a new laboratory test is needed for a better examination. One of the most recent quantitative methods that have already caught the attention of many researchers is the detection of bacteria by Real Time PCR method. The aim of this study is to identify bacteria of the genus Brucella, both in milk and cheese samples, by the method of Real Time PCR. 25 samples of cow's milk and twenty-five samples of cheese were collected from different parts of Shahrekord city. DNA extraction was performed using the DNA extraction kit (Cinnagen company) for the molecular diagnosis. Then, by the use of Real Time PCR reaction (Corbett Rotor-Gene Model, manufactured in Australia), samples were studied. In this study, fifty samples were examined, and only two samples (4%) were diagnosed with Brucella abortus, meanwhile, there were no reports on infections by Brucella in the cheese samples. Conclusion This study shows that molecular techniques such as Real Time PCR can be used as a complementary method for the detection of Brucella, alongside all the other common methods.

Pancreatic cancer is one of the most deadly and aggressive cancers; Fluorouracil induces apoptosis and cell cycle arrest in cancer cells. In the present study; the effect of Fluorouracil on different stages of the cell cycle and the expression of genes involved in the internal pathway of apoptosis in the AsPC-1 cell line (human pancreatic cancer) were investigated. In order to do so, MTT assay was used to evaluate the cytotoxic effect of Fluorouracil on AsPC-1 cell proliferation; The type of induced cell death and cell cycle changes were investigated by flow cytometry; changes in the expression level of genes (BAX, Bcl-2, APAF-1, Caspase-3, Caspase-9, p53, p21) were examined by Real-time PCR. Quantitative data were analyzed at the significant level of (p<0.05). The MTT assay results showed that Fluorouracil decreased AsPC-1 cell proliferation in a concentration-dependent manner. The results of flow cytometry analysis showed that increased percentage of apoptotic cells in the treated cells; Fluorouracil induces S phase cell cycle arrest in AsPC-1 cells and reduced distribution in the G1 phase. The Real-time PCR results in treated cells showed an increase in the expression of genes in the mitochondrial apoptotic pathway as well as genes effective in regulating the cell cycle. Fluorouracil reduces cell proliferation and induces apoptosis by increasing the expression of genes involved in the Intrinsic apoptotic pathway in AsPC-1 cells; Fluorouracil also caused cell cycle arrest in these cells by regulating the (p53, p21) genes.

Glioblastoma is an aggressive malignant tumor of the brain and spinal cord. Several lines of evidence indicate an important oncogenic role of highly non-coding RNA (lncRNA) molecules in a variety of cancers including glioma, however, the tumorigenic function of lens in glioma remains largely unclear. Increasing evidence has shown that lncRNAs, including LncRNA SNHG15, play an important role in the pathophysiology of human diseases, especially in the pathogenesis and progression of cancers. In this study, 25 paraffin tissue blocks of patients with glioblastoma multiforme and tumor margin tissue were collected and RNA extracted. Synthesis of cDNA and analysis of SNHG15 expression was done by Real-Time PCR technique. ROC curve was drawn to check the biomarker value and statistical analysis was done by GraphPad Prism v.8.0.1. An increase in the expression of SNHG15 was recorded in the tissue samples of patients compared to the tissue of the tumor margin (p < 0.0001). There was a significant relationship in the expression of this gene in patients aged more than 50 years and less than 50 years (p = 0.6573), Frontal, Temporal, Parietal, Occipital, and Other locations (P value = 0.9802), survival in more than 12 months and less than 12 months (p = 0.5007) was not observed, only in male and female sex (p = 0.0001) was registered. The expression of LncRNA SNHG15 was recorded in the tumor tissue of patients with glioblastoma multiforme more than in the peripheral tumor tissue. By examining the ROC curve, it is possible that LncRNA SNHG15 can be proposed as a biomarker, but it needs more studies.

Enteroviruses (E.V.s) and human parechovirus (HPeV) play an important role in childhood diseases, from mild infections to severe illnesses which cause morbidity and mortality. Since the diagnostic procedures are not routinely accessible in the clinical laboratory, the epidemiology of enterovirus and parechovirus remains ambiguous. The current study investigates the frequency of human Enteroviruses and Parechovirus in hospitalized children.

Nasopharyngeal swabs were collected from children younger than five years with severe acute respiratory symptoms admitted to the hospital and screened for E.V.s and HPeV using RT-PCR and a two-step real-time PCR.

From 80 nasopharyngeal swabs, viral agents were detected in 4 (5%) hospitalized children with ARTIs. All of these positive cases were infants. The results of RT-PCR of pan-HPeV demonstrate no detection of HPeV. Four samples were positive for enterovirus (5%). The frequency of E.V.s and HPeV between different sexes was not significant. Also, the distribution of E.V.s and HPeV among the other age groups was not significant (P>0.05).

Enteroviruses and human Parechoviruses can cause severe consequences like neurological impairments, and more importantly, the epidemiological characteristics information in this regard is limited. Therefore, further research on viral respiratory tract infections is needed to improve public health and prevent severe complications.

Enterococcus faecalis species are significant issue in severe nosocomial infections. In this study we aimed to investigate the effect of ionic liquids based on amino acids on pathogenic genes of this bacterium using Real time PCR.

In this study, 100 samples of oral infections were taken from patients and Enterococcus faecalis strains were identified using biochemical tests. The presence and frequency of cyl, hyl and esp genes and their expression under the influence of ionic fluid were evaluated using real time-PCR.

From the total samples taken, twelve Enterococcus faecalis bacteria were isolated and identified. The highest frequency of pathogenic genes was related to cyl gene which was present in 11 strains and the expression of target genes in the treated group under the influence of ionic fluid decreased compared to the control group (p <0.05). With decreasing ion-based amino acid concentration, the ability of bacteria to form biofilms increased (p <0.05).

Widespread antibiotic resistance is the most important concern about Enterococcus and cytolysin is a gelatinase enzyme and one of the most important invasive agents of Enterococcus faecalis. The results of this study provide a basis for promoting the use of new antibacterial agents, including ionic fluid, to prevent further pathogenicity of Enterococcus faecalis.

Sargassum ilicifolium is a multicellular brown algae that contains beta - carotene and fucoxanthin pigments, soluble acids of immune stimulants such as phycocyanin, polysaccharide, iron, zinc and essential amino acids, which promote safety. In this study, the effect of Sargassum iliisifolium algae extract on linear wound healing percentage and its effect on TGF β 1 and VEGFA gene expression in rats were investigated.

This research is an experimental study that was performed in the laboratory of Parand University of Medical Sciences. For this study, 30 male Wistar rats weighing 180 gr were used and 2 cm deep linear dermis was created from the neck to the waist of rats. Mice were divided into 3 groups of ten: The control group,The group received alpha ointment as a positive control drug and The group algae extract of Sargassum iliisifolium. The groups were examined for both wound size and expression of VEGFA and TGFB1 genes .

The alpha group had a 10.38 fold increase in TGFΒ1 gene expression compared to the control group, with a probability value of P <0.001. Compared with the algal group and the control group, it had a 5.39 increase in gene expression with a probability equal to P <0.001 . Also in assessing VGEFA gene expression :the alpha group significantly increased from the control group by 1.86 times the increase in gene expression with a probability value of p <0.044 , the sargassum algae group compared to the control group by 1.70 There is significant difference between the increase in expression and the probability value of p <0.040 . In terms of the percentage of wound healing, it shows that: the control group, reduced the wound length from 2 cm to 0.64 cm, the alpha group reduced the wound length from 2 cm to 0.5 cm and sargassum algae, the amount of wound has been reduced from 2 cm to 0.3 cm

Sargassum iliisifolium algae extract due to its antibacterial, anti -inflammatory, antioxidant and antimicrobial properties and due to the presence of alginate in it biometrically heals wounds. Available in the market, however, sargassum algae extract make a significant difference in increasing the expression of TGFβ1 and VEFGA gene .sargasum algae extract may use as a wound healer to heal skin damages

One of the most malignant and common brain tumors is glioblastoma multiforme (GBM). Patient survival is relatively poor, and due to the difficulty of early diagnosis, most patients die within one year of diagnosis. The aim of this study was to investigate the expression of YKL39 as a possible biomarker in glioblastoma multiforme and to evaluate the miR of this gene.

In this study, 25 tissue samples with glioblastoma multiforme before treatment and 25 samples of healthy tissue with glioblastoma multiforme tumor were collected as a control sample. RNA extraction and cDNA synthesis were performed. YKL39 gene expression was assessed using real-time PCR and ACTB gene was used as internal control. Statistical analysis of data was performed using GraphPad Prism software version 8. The Roc curve was used to evaluate the biomarker value of YKL39 gene.

High expression of YKL39 (P **** <0.0001) was observed in glioblastoma tissue samples. Expression showed that YKL39 gene was not statistically significant with the age of patients with P value = 0.2971 and gender with P value = 0.888.

The increase in YKL39 gene expression in glioblastoma tissue samples is significant compared to healthy tumor margin tissue. The Roc curve showed that the YKL39 gene could not have biomarker value.