جستجوی مقالات مرتبط با کلیدواژه « Gene expression » در نشریات گروه « پزشکی »

Andrographolide has been studied on different types of human cancer cells, but very few studies have been conducted on oral cancer. The study aimed to evaluate the anticancer potential of Andrographolide on an oral cancer cell line (KB) through in-silico network analysis and in vitro assays.

The in-silico analysis involved the determination of drug-likeness prediction, prediction of common targets between oral cancer and andrographolide, Protein-Protein Interactions (PPI), hub genes, top 10 associated pathways by Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway, Gene Ontology (GO), and molecular docking experiments. In vitro assays comprised MTT assay, apoptosis assay, cell cycle analysis, intracellular reactive oxygen species (ROS) measurement, mitochondrial membrane potential (MMP), anti-migration activity, and gene expressions using Polymerase Chain Reaction (PCR).

Fifteen common genes were obtained and were seen to be involved in cellular proliferation, regulation of apoptosis, migration of cells, regulation of MAPK cascade, and regulation of cell cycle. The most common genes involved in the top 10 pathways were MAPK1, MAPK8, MAPK14, and IL6 which were seen to be associated with the MAPK signaling pathway which may be the key pathway through which andrographolide may aid in treating oral cancer. In vitro assays showed anti-proliferative properties, late apoptosis, and anti-migratory properties.

According to the results obtained, andrographolide has shown anticancer properties and has the potential to be used as a chemotherapeutic drug. The in-silico approach used in the present study can aid as a model for future research in developing efficient cancer treatments.

Prostate cancer is known as the second common cancer among men, and is introduced as the fifth cause of death. Apoptosis known as “Programmed Cell Death”, can be measured as a defense mechanism in response to cell damage, caused by diseases or exposure to toxic material, and has an important role in controlling cell population. For this, researchers are interested in studying cancer treatment for inducing apoptosis. Flaxseed, Linum usitassimum, has numerous benefits among other oilseeds. It contains a large amount of α-Linoleic Acid (ALA), dietary fiber, proteins, and Phytoestrogen.

The main goal of this research was to evaluate the expression of apoptotic genes induced with flaxseed extract in human prostate cancer cells in order to find out the cell death pathway induced by flaxseed extract.

In this study, PC-3 prostate cancer cells were cultured and the treatments were done using the flaxseed extract. The MTT test, IC50 calculations, Flowcytometry, and Real-Time PCR test for apoptotic genes were done and all the results underwent analysis.

The IC50 dose was obtained at 809 µg/ml (P < 0.05). Flowcytometry test was done for 600, 800, and 1000 µg/ml of extraction, and the result showed that most of the cells underwent necrosis. The Flowcytometry result was confirmed by qPCR test and showed the overexpression of TNF gene (P < 0.05).

The results showed the overexpression of all the studied genes after treatment by flaxseed extract. The Flowcytometry result was confirmed by qPCR and showed that the TNF gene was overexpressed.

This study aimed to evaluate the expression of tcdA, tcdB, and binary toxin genes (cdtA and cdtB) by Real-Time PCR and molecular typing of Clostridioides difficile isolated from patient diarrhea samples from Hamadan Hospitals, west of Iran.

The concentration of C. difficile toxins (CDTs) is associated with the severity of the disease and the mortality rate. Measuring CDT levels could provide a reliable and objective means of determining the severity of C. difficile infection (CDI).

From November 2018 to September 2019, 130 diarrhea samples were collected from hospitalized patients in three hospitals in Hamadan. C. difficle isolates were detected by culture and PCR. The presence of the genes encoding the toxin was identified by PCR, whereas the measurement of toxin expression was conducted using a relative Real-Time PCR technique. Genetic linkage of the isolates was also assessed by Ribotyping and Repetitive Extragenic Palindromic (rep-PCR) methods.

Among 130 diarrhea samples, 16 (12.3%) were positive for C. difficile. Genes encoding cdtA and tcdB were detected in all isolates, and 8 (50%) and 6 (37.5%) isolates were positive for the cdtA and cdtB genes. Real-time PCR results showed different expression levels of the toxin genes. A significant increase in the expression of the tcdA gene was observed compared with the control strain (P<0.05). Besides, more expression of cdtA gene was observed in the strains compared with cdtB gene. Ribotyping and rep-PCR results showed high genetic diversity of C. difficile among hospitals investigated.

We encountered toxigenic C. difficile strains with various toxin expression levels, ribotypes, and rep types based on the findings of this study. This indicated that various clones from various sources circulate in the hospitals and among patients.

The consumption of contaminated poultry meat is considered as a significant route of campy- lobacteriosis transmission. Lactic acid is a disinfectant agent with bactericidal effects on Campylobacter spp. The purpose of this study was to assess the low concentrations of lactic acid effect and different temperatures on the transcriptomic responses of Campylobacter jejuni (C. jejuni) adhesion and virulence-associated genes including peb4, ciaB, cdtA, cdtB, and cdtC.

The samples were incubated at 10°C and 22°C for 48 h upon exposure to 30% and 60% lactic acid. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of lactic acid was also determined. Then, gene expression was assessed using real-time polymerase chain reaction (RT-PCR).

Lactic acid had lower MIC and MBC levels at lower temperature. The utilization of both levels of lactic acid signifi- cantly reduced the expression of peb4, ciaB, cdtB, and cdtC genes over 48 h of incubation at 22°C. However, no significant difference was found in the expression of the cdtA gene between 10 and 22°C at 30% lactic acid.

These results highlight the potential of low-concentration lactic acid in the downregulation of adhesion and virulence-associated genes as well as reduction of C. jejuni pathogenicity.

Colorectal cancer (CRC) is a heterogeneous disease of the colon or rectum arising from adenoma precursors and serrated polyps. Recently, probiotics have been proposed as an effective and potential therapeutic approach for CRC prevention and treatment. Probiotics have been shown to alleviate inflammation by restoring the integrity of the mucosal barrier and impeding cancer progression.

In this study, we aimed to investigate the immunomodulatory effects of live and UV-killed Bacillus subtilis natto on the inflammatory response in CRC. Caco-2 cells were exposed to various concentrations of live and UV- killed B. subtilis natto, and cell viability was assessed using MTT assay. Gene expression analysis of IL-10, TGF-β, TLR2 and TLR4 was performed using RT-qPCR.

Our findings showed that both live and UV-killed B. subtilis natto caused significant reduction in inflammatory response by decreasing the gene expression of TLR2 and TLR4, and enhancing the gene expression of IL-10 and TGF-β in Caco-2 cells as compared to control group.

The results of this study suggest that live and UV-killed B. subtilis natto may hold potential as a therapeutic supplement for modulating inflammation in CRC.

This in silicostudy investigated the association between the local biosynthesis of cholesterol and mammographic density, the major risk of developing breast cancer, as a function of the three cellular components of breast tissue (epithelium, fatty, and non-fatty stroma).

The study compared the expression of 7 genes (HMGCR, FDPS, FDFT1, GGPS1, SQLE, LSS, and SREBF2) involved in the de novocholesterol biosynthesis, first, according to the radiological density (dense vs. non-dense breast) and, then, according to the cellular components of breast tissue, regardless of the radiological classification.

HMGCR, SQLE, and SBREF2 were significantly more frequently expressed in radiologically dense than in non-dense breasts (-1.70 vs. -1.41, P=0.0028; -1.20 vs. -1.11, P=0.0501; -3.63 vs. -3.31 P=0.0003; -0.92 vs. -0.76, P=0.0271, respectively). When the samples were reclassified based on their cellular components as highly fatty and highly non-fatty, HMGCR, SQLE, and SBREF2 were significantly more frequently expressed in highly non-fatty samples (-1.48 vs. -1.94, P<0.0001; -3.39 vs. -4.18, P<0.0001; -0.77 vs. -0.94, P=0.0103, respectively), whereas LSS was overexpressed in high fatty ones (0.28 vs. -0.60, P<0.0001). Besides, while in the highly non-fatty subgroup, SREBF2 was positively associated with both HMGCR (r=0.53, P<0.0001) and SQLE (r=0.73, P<0.0001), in the highly fatty subgroup, these positive correlations disappeared (SREBF2*HMGCR: r=-0.19, P=0.3026) or substantially decreased (SREBF2*SQLE: r=0.41, P=0.0173).

Findings provide a compelling biological explanation for the clinical evidence that women with radiologically dense breasts are at a higher risk of developing cancer compared to those with non-dense breasts because of the prevalence of non-fatty tissue, where the altered expression of genes leading to an increased cholesterol production can contribute to the transformation of epithelial cells, and support the use of mammographic density as a reliable surrogate marker to identify women who may benefit from a preventive treatment aimed at reducing cholesterol production.

Given the cytotoxicity of chemotherapy drugs used in cancer treatment, there is a need to develop alternative agents to protect female fertility. This study investigated the effect of Actaea racemosa (A. racemosa) extract on mice ovarian cells and the damage caused by doxorubicin (DOX) to the mice ovaries.

We evaluated the effects of A. racemosa extract on mice ovaries (n=42) after DOX treatment. The mice were pre-treated with saline solution (controls) or with 0.5 or 5 mg/kg A. Racemosa extract. Afterward, during a period of 10 days, they were treated daily with one of the six protocols: (i) saline solution (control), (ii) 10 mg/kg DOX, (iii) 0.5 mg/kg A. racemosa extract, (iv) both DOX and 5 mg/kg A. racemosa extract, (v) A. racemosa extract (5 mg/kg), and (vi) both DOX and 0.5 mg/kg A. racemosa extract. At the end of these treatments, the ovaries were fixed for histopathological examinations. Ovarian follicular morphology, stromal cell density, collagen fibers, and TNF-α expression were evaluated. Some ovaries were fixed for transmission electron microscopy or stored at -80oC to study the mRNA expression for Caspase-3 and TNF-α.

The Mice treated with A. racemosa extract had reduced follicular degeneration and cell death after exposure to DOX. Ovaries of mice treated with 0.5 mg/kg A. racemosa extract had granulosa cells and oocytes with preserved ultrastructure, decreased immunostaining for TNF-α, and reduced Caspase-3 mRNA.

The A. racemosa extract supported follicular survival and protected the ovarian follicles and stromal cells against DOX-induced cytotoxicity.

 Cancer, the second leading cause of mortality worldwide, represents a global health challenge, primarily due to drug resistance. Vinorelbine is a chemotherapeutic agent that disrupts cancer cell growth by targeting microtubules and inducing apoptosis. However, drug resistance remains a formidable obstacle. This resistance is caused by various factors including genetic mutations, drug efflux mechanisms, and DNA repair systems. Resolution of this challenge requires an innovative approach. This study investigated the potential of small interfering RNA (siRNA) to target and downregulate a vinorelbine-resistant MCF-7/ADR breast cancer cell line.

 Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) 10% fetal bovine serum/penicillin/streptomycin. An siRNA targeting ABCB1 was designed and synthesized, and the cells were transfected with siRNA at final concentrations of 10, 20, and 30 nM. The3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to assess cell viability. ABCB1 mRNA expression levels were determined by real-time polymerase chain reaction (PCR).

 MCF-7 cells exhibited a higher sensitivity to vinorelbine than MCF-7/ADR cells. MCF-7/ADR cells exhibited resistance to vinorelbine at concentrations, 12.50 and 25.00 μM. Treatment with siRNA significantly reduced ABCB1 expression by 2.93-fold (P=0.0001). Similarly, co-treatment with siRNA and vinorelbine produced a substantial 2.89-fold decrease in ABCB1 gene expression in MCF-7 cells compared to that in MCF-7/ADR cells (P=0.0001).

 The results of the present study indicate that the concurrent use of siRNA and vinorelbine holds substantial promise as a therapeutic approach to overcome ABCB1-mediated multidrug resistance (MDR) in breast cancer. It is necessary to conduct comprehensive clinical trials to determine the true effectiveness of this combination therapy.

Pseudomonas aeruginosa, drug-resistant, causes health infections. Resistance to the preferred therapy meropenem is a serious threat. This study aimed to analyze changes in meropenem minimum inhibitory concentra- tion (MIC), changes in ampC, mexA, and oprD gene expression, and the correlation between MIC and ampC, mexA, and oprD gene expression after meropenem exposure.

Ten isolates of P. aeruginosa from the Clinical Microbiology Department, Faculty of Medicine, Universitas Indonesia were used. After the bacteria were shown to be sensitive to meropenem phenotypically, intrinsic resis- tance genes were detected using PCR. After meropenem exposure on Days 5 and 12, sensitivity testing was carried out with the concentration gradient method and RNA was detected using real-time RT-PCR.

All P. aeruginosa isolates that were phenotypically sensitive to meropenem had the ampC, mexA, and oprD genes. An increase in MIC, an increase in ampC and mexA gene expression, and a decrease in oprD gene expression were observed after meropenem exposure. There was a very strong and significant correlation (p ≤ 0.05) between MIC and oprD gene ex- pression after Day 12 of meropenem exposure.

Although there were no significant differences in MIC and ampC, mexA, and oprD gene expression between Day 5 and Day 12, there was a very strong and significant correlation between MIC and oprD gene expression on Day 12 (p ≤ 0.05). This indicates that decreasing oprD gene expression has the potential to increase meropenem resistance in Pseudo- monas aeruginosa.

Enterococcus faecalis is known as common pathogen for endodontic infections and cause secondary and refractory pulp periapical periodontitis. The bacteria can opportunistically colonize periodontal pockets and presents a possibility of infection developing in other organs. This research will investigate the dissemination of E. faecalis from the gingival tissue to the heart and kidney.

Three groups were formed, consisting of twelve male Sprague Dawley rats: a control group des- ignated as 0-day, and experimental groups labeled as 7-days and 14-days. Periodontitis induced by concurrent infection with sterile wire 0.2 mm insertion and E. faecalis inoculation is performed into the gingival sulcus located between the maxillary right 1st and 2nd molar teeth area. After euthanasia, tissue samples around the maxillary gingiva, maxillary jaw samples, kid- ney and heart tissues were obtained for quantitative Real-Time PCR assay and histopathological analysis.

Results showed at 7-days, there was an upregulation of E. faecalis gene expression in the gingiva, heart, and kidney samples as well as infiltration of the inflammatory cells at 7-days post induction, which consequently decreased at 14-days.

Thus, the study suggests dissemination of E. faecalis from gingival tissue to the heart, kidney which could be probable link between periodontal disease, heart, and kidney disease.

Sports activity increases PGC1α and Nrf2, the regulatory factors of mitochondrial biogenesis. This paper aims to study the impact of two-month sodium citrate supplementation with Moderate-Intensity Continuous Training (MICT) on PGC-1α and Nrf2 expression in diabetic rats.

Forty-five three-month-old male Wistar rats were haphazardly assigned to one of five equal groups (N=9): (1) healthy; (2) diabetic; (3) diabetes + exercise (DE); (4) diabetes+ supplementation (DS); and (5) diabetic + exercise + supplementation (DSE), matched according to their weights. After induction, exercises began on a treadmill for 8 weeks, five days a week. The MICT protocol ran at 70% of their maximum speed for 36 minutes. The rats supplemented with sodium-citrate- at 15 mmol/L in drinking water for two months. PGC-1α and Nrf2 expression were measured through Western blotting in the soleus muscle. Data were analyzed using univariate analysis of variance (ANOVA) and the Tukey post-hoc test. Cohen’s D effect size (ES) was calculated to compare the groups.

The results showed that induction of diabetes significantly reduced the expression of PGC-1α (P< 0.001; ES=1.36) and Nrf2 (P<0.088; ES=0.24), while exercise increased PGC-1α expression (P<0.001; ES=0.68). Sodium citrate supplementation, either alone or in combination with MICT activity, did not show a clear advantage for Nrf2 expression.

MICT activity and sodium citrate supplementation, by increasing PGC-1α expression, can be considered therapeutic strategies for diabetic patients. However, to increase Nrf2 expression, further studies with different exercise intensities and doses of sodium citrate supplementation are needed.

Ankylosing spondylitis (AS) is a chronic autoimmune disorder characterized by the fusion of vertebral joints and axial arthritis. The programmed death-1 (PD-1) inhibitory receptor has a pivotal role in controlling T cell function and may have a significant impact on the pathogenesis of autoimmune diseases such as AS pathogenesis.

To investigate PD-1 gene expression and its epigenetic regulation by detecting methylated CpG islands in the regulatory sites of the gene. This will provide insight into the mechanisms involved in the disease.

30 AS patients and 30 healthy individuals were examined to detect the 16 CpG islands in intron 1 using bisulfite conversion and methylation-specific PCR technique. In addition, RNA samples were isolated from fresh peripheral blood mononuclear cells (PBMCs), and after complementary DNA (cDNA) synthesis, the expression level of the PD-1 gene was evaluated using Real-Time PCR.

The CpG islands located in the intronic zone of the PD-1 gene were hyper-methylated in both the patients with AS and the healthy controls. The gene expression of PD-1 was significantly downregulated in AS patients compared with the controls (p=0.017). A negative correlation between the Bath Ankylosing Spondylitis Disease Activity Index and PD-1 gene expression was also revealed.

The low level of PD-1 gene expression is implicated in the pathogenesis of AS. However, in both groups, the methylation level of the intron 1 CpG islands of the PD-1 gene suggests that other regulatory mechanisms are more relevant to PD-1 gene expression than methylation in the intron.

HSV-1 is known as a very contagious virus and the main cause of cold sores or fever blisters. Herein, the aqueous extract of Areca catechu L. was evaluated for its anti-HSV-1 activity, compared to the standard control (acyclovir). Also, the effect of extract on the expression of UL46 and US6 genes that accumulate late in viral infection, was studied.

The aqueous extract was obtained by the maceration of powdered plant in boiling water. Its antiviral activity was evaluated on Vero cells infected with HSV-1 at different times: 2 h pre-infection, simultaneous infection, and 4 h post-infection, using MTT assay. The effect of extract on the expression of genes was investigated with quantitative real-time PCR.

The aqueous extract of A. catechu induced the inhibition of infection with the IC50 value of 110.52 ± 1.36 μg/ml. Also, it reduced the expression of UL46 when it was added 2 h pre-infection at 100 μg/ml. Moreover, reduction of expression of US6 was observed at the same concentration when the extract was used simultaneously with the occurrence of infection and 4 h post-infection.

A. catechu can be considered an essential element of natural-based anti-HSV-1 agents.