فصلنامه تازه های بیوتکنولوژی سلولی مولکولی
پیاپی 52 (پاییز 1402)

Rheumatoid arthritis(RA) is a complex and chronic disease of joint inflammation. Environmental factors that are important in genetically predisposed people are involved in the occurrence of the disease and its progress. This disease has a lot to related to genetics, including human class 2 antigens. In various studies, this relation is about 60%. This relationship is more positive in people with Anti-citrullinated protein antibodies (ACPA) than negative ones. The aim of this study is to investigate the relationship between cytokines and soluble proteins in the body of a patient with Rheumatoid arthritis before symptoms appear.

In this research, analysis with Enrichr, WebGestalt, and GeneMANIA was used to find genes related to the disease. In order to check the amount of cytokines predicted, ELISA and Real time RT-PCR methods were applied.

By examining the System Biology methods, the candidate genes included IL-6R, IL-6 and IL-23A. The samples were taken from 46 RA patients referred to the rheumatology department of the Babol hospital, and a significant decrease (Pv=0.1255) of IL-6 was observed in patients undergoing treatment and a significant increase (Pv=0.4414) in the amount of this cytokine receptor in the serum compared to healthy individuals. While the expression of IL6 and IL6R genes increased in the patient group compared to the healthy group.

Cytokine IL-6 as an inflammatory cytokine in treated patients decreased in serum and showed a significant increase in RNA level.

The process of obesity is complex and many factors, including two key hormones, leptin and insulin are involved. The homeostatic role of insulin in energy stability is reduced by insulin resistance. It is possible that leptin and insulin have a regulatory role on each other, therefore, the present study investigated the relationship between BMI, leptin , insulin and insulin resistance.

In this cross-sectional study, 140 women were examined in 2 equal groups with normal (18.5-24.9) or obese (≥ 30) BMI. After fasting for 10-12 hours, 10 cc of venous blood was taken from the subjects, Blood sugar was measured by glucose oxidase, insulin and leptin by ELISA method, and HOMA-IR index was calculated. The data were expressed as mean ± standard deviation. P < 0.05 was considered statistically significant.

There was a significant difference between the groups in terms of leptin level, and there was a significant correlation between BMI and plasma leptin level of the studied subjects, r=0.07 and P<0.01. Also, there was a significant relationship between serum leptin and fasting insulin (r = 0.05 and P < 0.01. In terms of the HOMA-IR, a significant difference between the groups was observed which was higher in obese than in normal.

BMI is the main factor in regulating leptin levels. Probably the continuity of insulin and leptin levels is due to the determining role of BMI on both of them.

The AKR1C3 protein plays a very important role in various diseases, especially cancer, and is an attractive target for drug development in hormone-dependent cancers and inflammatory diseases. Therefore, the aim of the present study was to investigate the inhibitory mechanism of AKR1C3 protein and to design new inhibitors.3زز

In the present study, the three-dimensional structure of AKR1C3 was obtained from the three-dimensional protein structure database using molecular systems. Then, the obtained structure was optimized using the molecular dynamics technique, and then the protein interactions with the existing inhibitors were investigated using the docking technique and Autodock4 software. Then, in order to perform molecular dynamics, AMBER software version 18 was used. Finally, 4 new inhibitor combinations with Data Warrior software were designed as new inhibitors and introduced to inhibit AKR1C3.

The amino acids tyrosine 55 and histidine 117 are involved in the formation of strong hydrogen bonds with the designed inhibitors. The amino acids tyrosine 55, histidine 117, and phenylalanine 311 play an important role in the interaction of proteins with designed inhibitors. The study of binding energy of these compounds to protein showed that the binding energy values ​​of these compounds with AKR1C3 protein are in the range of -17.8 to -29.1 kcal / mol, which can be well competitive with known protein inhibitors. Meanwhile, the structure of D1 as a designed compound with a binding energy of -32.9 kcal / mol was able to show the greatest tendency for protein among the inhibitory compounds known and designed in this study, which can be considered as a compound with a high potential for Introduce inhibition of AKR1C3 protein in laboratory and clinical studies.

The results of docking and molecular dynamics studies showed that the new compounds based on mefenamic acid could be potential compounds to inhibit the AKR1C3 protein, and by making appropriate changes in them, these target compounds could be used as effective compounds to inhibit and fight disease. Used inflammatory disease sentence.

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.

PD-L1, one of the most important immune-regulating molecules in all cancers including breast cancer, suppresses the immune system against tumor cells. MiRNAs control PD-L1 expression as therapeutic tools and targets. This study investigated the role of miR-200c in reducing PD-L1 expression and breast cancer cell proliferation.

In this study, miR-200c targeting the PD-L1 3'UTR was confirmed by bioinformatics and dual-luciferase assay. PD-L1 expression in breast cancer tissues and cell lines was checked compared to normal tissues and cell lines. qRT-PCR was used to measure PD-L1 expression after miR-200c transfection into breast cancer cell line. The MTT test was used to determine the effect of miR-200c introduction on cell proliferation. 

The 3'UTR region of the PD-L1 gene was identified as a target for miR-200c using bioinformatics software and a dual luciferase assay. The decrease in miR-200c expression in breast cancer cells is directly related to the increase in PD-L1 expression (r =-0.68, P value₌ 0.0158). Also, increasing the expression of miR-200c in MDA-MB231, BT549, and MCF7 breast cancer cells decreased the expression of PD-L1 by (0.217±2.3, 0.021±2.11, and 0.012±0.146, respectively). The increased expression of miR-200c decreased the proliferation rate in MDA-MB231 by 0.45 ± 0.012, BT549 by 0.256 ± 0.2, and in MCF7 by 0.164 ± 0.02, compared to non-transfected cells. (Mean ± SE)

MiR-200c may be able to prevent breast cancer progression by targeting the PD-1/PD-L1 pathway, reducing PD-L1 expression, and reducing breast cancer cell proliferation.

Harmal or Spand is one of the medicinal plants with the scientific name of peganum harmala and belongs to the Zygophylaceae family. Beta-carboline belongs to the group of indole alkaloids and is present in different parts of the plant, especially seeds and roots. Beta-carbolines exert a range of neurophysiological and toxic effects, including antidepressant activity, vasodilation, anti-cumulative effects of platelets, and effects on drug withdrawal and appetite. The aim of this study was to extract and measure beta-carbolines as well as phenolic compounds of Pecan plant species and to investigate its antioxidant properties.

For this purpose, this plant was collected from around Zahedan and methanolic extract of leaves and seeds were extracted by Soxhlet method. Then different phytochemical analyzes were performed on the leaves and seeds of this plant. Quantitative and qualitative analyzes included identification of anthocyanins, flavonoids, tannins, anthraquinones, antioxidant test of the extract, FTIR analysis of the extract, quantitative and qualitative HPLC analysis of the extract and alkaloids.

The results showed that Harmine was toxic at a much lower dose than the seed extract, and the seed extract can be toxic in low doses than leaf extract. In the leaf extract, compared to the seed extract, the amounts of alkaloids are small, and its antioxidant effects are greater due to the higher amounts of polyphenols and flavonoids.

The results showed that although the antioxidant properties of methanolic extract of leaves (due to the presence of flavonoids and phenolic compounds) are higher than seeds, but there is more beta-carboline in plant seeds.

Skin is a soft tissue that protects the whole body and internal tissues against external hazards such as burns or injuries. Risks cause loss of the skin's defense mechanism and severe infections and wounds that inhibit the healing. Therefore, it is very important to develop effective and fast skin repair. The aim of this study was to prepare a porous scaffold for wound covering made of Polycaprolactone (PCL), the core of this scaffold is made of Carbon Quantum Dot (CQDs), Iron nanoparticles (Fe), and Chitosan (Cs).

The composition of Fe-CQDs was synthesized by ammonium-hydrogen-citrate under hydrothermal conditions and Poly capro lactone/Chitosan/Carbon-quantum dot-iron scaffold by electrospinning method. The synthesized scaffolds were evaluated for morphology, functional groups, structure, and composition by Fourier-transform infrared spectroscopy (FTIR) , Scanning Electron Microscopy (SEM), and X-ray Powder Diffraction (XRD) analyses, respectively. After characterizing the fabricated scaffold, the viability of L929 fibroblast stem cells was obtained. 

SEM results showed successful integration of polycaprolactone scaffold with chitosan and carbon-iron quantum dot-iron. The average size of polycaprolactone/Chitosan/Carbon-quantum dot-iron nanocomposite was in the range of 32.6-135.0 nm. FTIR results showed the formation of bonds between carbonate and iron nanoparticles. A vibrating magnetometer (VSM) proved superparamagnetic of carbon-quantum dot-iron magnetic nanoparticles. At a concentration of 2% carbon-quantum dot-iron, the scaffold had good biocompatibility, which was proved by a cytotoxicity test.

The results showed that Polycaprolactone /Chitosan /Carbon-quantum dot-iron (PCL/Cs/ CQD-Fe) nanocomposite improves the wound healing process due to its non-toxicity. The biocompatible PCL/Cs/ CQD-Fe nanocomposite is a scaffold of interest in wound dressing that greatly accelerates the skin regeneration process.

One of the environmentally friendly synthesis methods is the synthesis of nanoparticles using plants. Silver nanoparticles are metal nanoparticles with anticancer activity. This study aims  to investigate the effect of silver nanoparticles synthesized from Digitalis nervosa hydroalcoholic extract on MCF-7 breast cancer cell line.

In this experimental study, silver nanoparticles were synthesized from the hydro alcoholic extract of the Digitalis nervosa.MCF-7 cells were then treated with different concentrations of silver(3.125, 6.25, 12.5,25,50 and 100 μg /ml).The toxicity effects of silver nanoparticles were analyzed by MTT method and after determining the concentration of 50% lethality of silver nanoparticles, the expression changes of P53,Bax, Bcl2 and CDH1 genes were investigated by Real - time PCR method. The obtained data were analyzed using SPSS software and one-way variance statistical test and Tukey's test.

The results of MTT showed that the effect of silver nanoparticles on the viability of MCF-7 cell line is dose-dependent and the highest cytotoxicity is related to the concentration of 100 and the lowest is related to 6.25 μg /ml in a significant way. (P<0.001) Also, the effect of silver nanoparticles with a concentration of 50% lethality (19.55 micrograms/ml) on the expression of P53, Bax, Bcl2 and CDH1 genes showed significant changes compared to the control gene.

Silver nanoparticles synthesized by green method from Digitalis nervosa  induce apoptosis in MCF-7 cell line and the use of these nanoparticles can be considered as a promising choice in the treatment of breast cancer.