Hepatocellular carcinoma (HCC) is one of the most aggressive cancer. MicroRNAs (miRNAs), are small noncoding regulatory RNAs, that function post-transcriptionally. MiRNA deregulation was observed in the development and progression of HCC. In this study we aimed to investigate the expression levels of four miRNA (mir-33a, mir-203b, mir361-3p and mir-424) in HCC patients in comparison to healthy individuals. Venous blood samples were collected from both HCC patients and healthy individuals. In order to determine relative expression levels of mir-33a, mir-203b, mir361-3p and hsa-mir-424 in HCC patients using probe-based Quantitative Real Time PCR (qRT-PCR) is performed. The cycle threshold (Ct) results were analyzed according to the 2???Ct method and statistical analyses was performed by IBM SPSS Statistics for Windows, version 15. qRT-PCR analysis revealed that the expression level of mir-33a (fold change: 7.3 and p<0,001), mir-203b (fold change: 4.6 and p<0,001), mir361-3p (fold change: 5.1 and p<0,001) were down regulated compared to healthy individuals and mir-424 did not show any significant change between the HCC patients and controls. Our results indicated that mir-33a, mir-203b and mir-361-3p, may significantly contribute to tumor pathogenesis in HCC and would have potential to be used as a non-invasive biomarker for cancer therapy.

The aerial parts of Satureja metastasiantha were hydrodistilled for 3 h using a Clevenger-type apparatus. The essential oils were analyzed by gas chromatography/flame ionization detector and gas chromatography/mass spectrometry, simultaneously, the main compounds of which were characterized as p-cymene (22.3%), thymol (21.0%), carvacrol (18.4%), and γ-terpinene (12.1%). Antioxidant capacity, acetylcholinesterase and butyrylcholinesterase inhibition effects, and antimicrobial and antifungal properties of the species were evaluated. The anticholinesterase activity of the essential oil of S. metastasiantha was observed with 30% inhibition at 200 μg/mL. The essential oil of the species showed activity against Staphylococcus aureus with 128 μg/mL minimum inhibitory concentration value.

Hepatocellular carcinoma is the most common primary malignant liver tumor. Mitochondrial DNA copy number has been shown to be associated with various malignancies. However, there has not been any study on the absolute quantification of mtDNA copy number in hepatocellular carcinoma. The aim of this study was to develop a new method for absolute quantification of mtDNA copy number and to relatively quantify the variations in the mtDNA copy number in hepatocellular carcinoma patients in comparison with healthy individuals. Venous blood samples were collected from both hepatocellular carcinoma patients (34) and healthy individuals (34). Circulating cell-free DNAs were isolated and the relative quantification of mtDNA copy number variation was determined using quantitative polymerase chain reaction and digital polymerase chain reaction. It was found that the relative mtDNA copy number was significantly decreased in hepatocellular carcinoma patients in comparison with the control group (p<0.05). The median (range) and average of relative mtDNA/β-actin gene of the patients were determined as 42.8 cp/μL (11.1–88.5) and 45.1 cp/μL, respectively, while the median (range) and average relative mtDNA/β-actin gene of the control group were determined as 102.8 cp/μL (55.1–291.8) and 138.7 cp/μL, respectively (p<0.05). When quantitative polymerase chain reaction and digital polymerase chain reaction were compared, mtDNA/β-actin gene copy number ratio of digital polymerase chain reaction results was found to be 1.76-fold more than that of quantitative polymerase chain reaction results. Circulating mtDNA copy number was decreased in hepatocellular carcinoma patients in comparison with healthy individuals, and we suggest that it can be used as a noninvasive biomarker for hepatocellular carcinoma diagnosis in the future.

Objectives: Cell-free DNA (cfDNA) is a promising biomarker for cancer diagnostics. Blood is used as a liquid biopsy material which transports many types of biological components such as protein, circulating tumor cells, cfDNA, and extracellular RNA molecules. In this study, the yield and purity of extracted DNA were compared of seven different in-house DNA isolation methods and two different commercial kits from human blood samples. Methods: Different in-house methods, guanidine thiocyanate (GuSCN) with silica method, phenol-chloroform-isoamylalcohol method-1, phenol-chloroform-isoamylalcohol method-2, phenol-chloroform-isoamylalcohol with ammonium acetate method, sodium N-lauryl sarcosine with glycogen method, sodium acetate method, sodium chloride with silica method and two different commercial kits “Quick-cfDNA serum & plasma kit” and “plasma/serum cell-free circulating DNA purification mini kit” were used. After DNA isolation, methods were compared with real time PCR technique (qPCR) and spectrophotometric methods. Results: The results showed that the newly modified GuSCN with silica method gave much higher purity and yield than any other analyzed in-house isolation methods and commercial kits that were used. Conclusions: The GuSCN with silica method is simple to set up, easy to use, inexpensive and environmentally safe. The method would be an alternative method for cfDNA isolation from liquid biopsy samples.