Cell and Tissue Journal

Abstract Introduction:Cartilages are inherently avascular and have limited repair capacity after injury. If not treated properly, these injuries can lead to progressive joint destruction and diseases such as osteoarthritis. Current treatment methods, including surgery and cartilage transplantation, face limitations such as inadequate access to healthy tissue, graft rejection, and poor-quality cartilage formation. Therefore, finding new strategies to stimulate efficient cartilage regeneration has become a critical need in regenerative medicine.In recent years, exosomes derived from mesenchymal stem cells (MSCs) have emerged as a promising tool in tissue regeneration. These extracellular vesicles contain growth factors, miRNAs, and proteins involved in tissue repair that can transmit therapeutic signals to target cells. Glucosamine, a naturally occurring amino monosaccharide, is an important precursor in the synthesis of glycosaminoglycans (GAGs) and proteoglycans, which are major components of the extracellular matrix of cartilage. This compound is widely used as a dietary supplement or drug in the treatment of degenerative joint diseases such as osteoarthritis. As a common supplement in the treatment of joint diseases, glucosamine plays an important role in stimulating the synthesis of cartilage extracellular matrix, promoting the differentiation of stem cells into chondrocytes, and inhibiting cartilage destruction.


Aims: The aim of this study was to investigate the combined effect of mouse bone marrow stem cell-derived exosomes and glucosamine on the expression of cartilage-specific genes including Sox9, Acan, Col2a1, and Col10a1.
Materials and methods: Bone marrow mesenchymal stem cells were prepared from NMRI mice. The mice were euthanized by cervical dislocation, the femoral heads were removed, and the bone marrow contents were transferred into a cell culture flask using a syringe containing culture medium. The bone marrow cells were cultured and were ready for use after 3 to 5 passages. The cell supernatant was separated, and exosomes were extracted from the cell supernatant by successive rounds of centrifugation followed by ultracentrifugation. Exosomes were characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM), transmission electron microscopy (TEM), and dynamic light scattering (DLS). The effects of exosomes and glucosamine on the expression of Sox9, Acan, Col2a1 and Col10a1 genes in mesenchymal stem cells were investigated in the presence of chondrogenic medium.
Results: The identification results from multiple characterization techniques confirmed the presence of exosomes with a size of approximately 100 nm. Real-time PCR results showed that the expression of Sox9, Acan and Col2a1 genes in stem cells treated with a combination of exosomes and glucosamine increased significantly compared to the other groups after 14 days of treatment, while the expression of the Col10a1 gene decreased significantly compared to the other groups.
Conclusion: This study demonstrated that the combination of exosomes derived from bone marrow mesenchymal stem cells and glucosamine can synergistically increase the expression of key genes in chondrogenic differentiation (Sox9, Acan and Col2a1) while reducing the expression of the Col10a1 gene (related to chondrocyte hypertrophy). These findings suggest the therapeutic potential of this combination in stimulating cartilage regeneration and preventing further degradation. Given these promising results, this approach should be further investigated in clinical studies as a novel therapeutic strategy for cartilage damage.

Abstract Cancer remains a major global health challenge, with breast cancer incidence steadily increasing in women. Its development involves uncontrolled cell proliferation and the ability of cancer cells to invade healthy tissues, forming tumors and metastases. Conventional therapies, including surgery, chemotherapy, and radiotherapy, face limitations such as damage to normal cells, systemic toxicity, and drug resistance. These challenges have spurred the development of targeted drug delivery systems, with nanoparticles emerging as a promising solution. Nanoparticles, due to their small size, high surface-to-volume ratio, and modifiable surfaces, can efficiently deliver therapeutic agents to target cells while reducing systemic exposure. Iron oxide (Fe3O4) nanoparticles offer magnetic and chemical properties that enable tumor-specific guidance and imaging. Surface modification with polyethylene glycol (PEG) enhances nanoparticle stability, prolongs circulation, and provides attachment sites for targeting ligands such as folic acid and glucose. Polylactic acid (PLA), a biodegradable polymer, improves drug-loading capacity and allows controlled release. Combining PLA with PEG and targeting ligands produces multifunctional nanoparticles capable of selective cancer cell recognition, controlled drug release, and reduced effects on healthy tissues.

In this study, dual-ligand PLA-PEG nanoparticles functionalized with folic acid and glucose and incorporating Fe3O4 were developed for targeted delivery of curcumin to triple-negative breast cancer cells (Hs-578T). Nanoparticles PPF (PLA-PEG-FA), PPG (PLA-PEG-Glu), and PPGF (PLA-PEG-Glu/FA) were synthesized via reaction of PLA-acrylate with the respective NH2-PEG derivatives, followed by dialysis and freeze-drying. Curcumin and Fe3O4-OA were incorporated using sonication and emulsification in PVA, followed by solvent removal, washing, and filtration to remove unencapsulated drug and large aggregates. Structural and chemical characterization was performed using 1H-NMR and FTIR spectroscopy, morphology was analyzed with TEM, and size and zeta potential were determined by dynamic light scattering. Drug encapsulation efficiency was measured spectrophotometrically, and release kinetics were evaluated at pH 7.4 and pH 4.5 to mimic normal and tumor environments. Cytotoxicity was assessed using MTT assays, while apoptosis induction was analyzed via Annexin V/propidium iodide staining and flow cytometry.

Results demonstrated that PLA-PEG-based nanoparticles were biocompatible with Hs-578T cells, showing minimal toxicity. Curcumin encapsulation enabled controlled and sustained release, particularly in acidic conditions representative of tumor microenvironments. Nanoparticles containing both folic acid and PEG (PPGF) exhibited higher drug release than PEG-only nanoparticles, emphasizing the role of targeting ligands in facilitating drug delivery. Cytotoxicity assays revealed significant inhibition of Hs-578T cell proliferation, with dual-ligand nanoparticles showing the lowest IC50 values. Flow cytometry confirmed that apoptosis, rather than necrosis, was the primary mode of cell death, indicating targeted activation of programmed cell death pathways. This enhanced apoptotic effect likely results from increased cellular uptake and precise intracellular delivery of curcumin.

The study confirms that multifunctional PLA-PEG nanoparticles, incorporating Fe3O4 and targeting ligands, provide an effective platform for cancer-specific drug delivery. They combine biocompatibility, controlled release, tumor-specific accumulation, and enhanced induction of apoptosis, collectively improving therapeutic efficacy while minimizing systemic toxicity. The nanoparticles’ pH-sensitive release ensures minimal drug exposure to normal tissues, reducing side effects and potentially allowing lower therapeutic doses. The use of dual ligands, folic acid and glucose, further increases targeting precision and drug accumulation in cancer cells. These findings support the potential of multifunctional nanocarriers as a promising strategy for next-generation targeted cancer therapies, offering a safe, efficient, and versatile approach for improving treatment outcomes and overcoming limitations associated with conventional chemotherapy.

Abstract Introduction: Okra (Abelmoschus esculentus) is an annual culinary plant of Malvaceae family with high nutritional and medicinal values. This plant is widely cultivated in different regions of Iran as well as the world.
Aims This study investigated the fatty acid composition and antioxidant activity of seed oils extracted from seven okra cultivars, including: White, Sultani, Red, Velvet, Green, Texas, and Fawn. Material and methods: The mature and intact seeds of each cultivar were powdered and their oils were extracted using the modified Folch et al. method. The fatty acid methyl esters were obtained from seed oils using transesterification method. Then, the oils were injected to GC and GC/MS apparatus to detect their fatty acids profile. The DPPH method was applied to detect antioxidant activity of the extracted okra oils. Data was analyzed using SPSS and MVSP software. Results: The seed oils in all cultivars were comprised both saturated and unsaturated fatty acids, while, their total percentages varying among these cultivars. The highest total saturated fatty acid content (42.53%) was observed in the white cultivar, while the lowest (36.41%) was detected in the Sultani cultivar. Palmitic acid was the predominant saturated fatty acid in all the cultivars, with its highest (35.74%) and lowest (30.7%) percentages were found in the white and Texas cultivars, respectively. Stearic acid was the second most abundant saturated fatty acid in all examined oils, ranging from 3.69% (Sultani cultivar) to 4.94% (white cultivar). Some trace amount saturated fatty acids such as heptadecenoic, arachidic, behenic, and lignoceric acids were present in all the cultivars, with lauric acid had a restricted distribution and was only detected in a few cultivars, including: velvet, Texas, and white cultivars. Unsaturated fatty acids constituted 57.33% to 63.33% of the oils composition, with the lowest and highest percentages in white and Sultani cultivars, respectively. Linoleic acid was the primary unsaturated fatty acid in all the cultivars except white cultivar, where oleic acid was detected as predominated unsaturated fatty acid. The highest linoleic acid percentage (11.40%) was detected in red cultivar and the lowest (8.25%) in white cultivar. The amount of oleic acid ranged from 18.20% (red cultivar) to 49.30% (white cultivar). Some trace amount unsaturated fatty acids such as myristoleic, palmitoleic, trans-heptadecenoic, linolenic, gondoic, and erucic acids were present in all the cultivars. Additionally, all the cultivar had eicosadienoic acid in a trace amount, except for white cultivar. Phytochemical analysis using UPGMA dendrogram and PCA analysis divided these cultivars into two main clusters, confirming significant phytochemical diversity. It seems that change in the predominant unsaturated fatty acid, oleic acid in the white cultivar instead of linoleic acid. Additionally, the presence of palmitic acid as the dominant fatty acid in the velvet and white cultivars, were two main reasons for placement of white, green and velvet cultivars far from other cultivars. Antioxidant activity assessed by the DPPH method revealed IC50 values ranging from 689.23 µg/mL in green cultivar to 2551.12 µg/mL in Texas cultivar. Discussion: the seed oil profile of white cultivar is more different from the others. Moreover, the week or trace antioxidant activity was detected for the oils of diverse okra cultivars. These findings highlight the nutritional and industrial potential of okra seed oils. Additionally, the oil profile can strongly influence the antioxidant activity of the oils.

Abstract Introduction
Gene therapy involves transferring genetic material into target cells to correct mutations or introduce new biological functions. Among delivery systems, lentiviral vectors are considered efficient and reliable tools due to their ability to integrate stably into the host genome and transduce both dividing and non-dividing cells. This property provides long-term gene expression, which is highly valuable for therapeutic and experimental applications.
The PDX1 (Pancreatic and Duodenal Homeobox 1) gene plays a central role in pancreatic organogenesis and regulation of insulin-producing beta cells. It acts as a transcription factor controlling genes critical for endocrine differentiation and insulin secretion. Chick embryos are a useful experimental model due to their accessibility, rapid development, and the responsiveness of their fibroblast and germ cells to gene transfer systems. These features make them suitable for studying gene delivery efficiency and expression stability.
Aim
The aim of this study was to construct and produce recombinant lentiviral vectors carrying the PDX1 gene in HEK293T-LentiX cells and evaluate their transfer efficiency in chick embryonic fibroblast and germ cells. This work was conducted to assess the potential of lentiviral systems for stable gene delivery in avian cells.
Materials and Methods
HEK293T-LentiX cells were selected as producer cells due to their high transfection efficiency and viral packaging capability. They were cultured in DMEM supplemented with 10% fetal bovine serum, penicillin, and streptomycin under standard incubation conditions (37°C, 5% CO₂).
Lentiviral particles were generated using a three-plasmid packaging system, including a transfer vector containing the PDX1 gene and two helper plasmids. Transfection was carried out using the calcium phosphate method. After 48–72 hours, the viral-containing supernatant was collected, filtered, and concentrated.
Viral titers were determined by evaluating GFP expression in target cells through fluorescence microscopy and flow cytometry. Chick embryonic fibroblast and primordial germ cells were isolated and infected with various viral concentrations in the presence of 8 µg/mL polybrene to enhance infection. After incubation, cells were examined for GFP signal as evidence of successful gene transfer.
Results
High-titer recombinant lentiviral particles were successfully produced in HEK293T cells. Fluorescence microscopy revealed strong GFP expression, confirming the presence of functional viral particles. Flow cytometry analysis provided quantitative confirmation of high viral titers.
Following transduction, chick embryonic fibroblast and germ cells exhibited clear GFP expression, indicating efficient infection and gene transfer. The PDX1 gene was successfully delivered and expressed within target cells. Although transduction efficiency varied slightly between cell types, the overall results demonstrated that the lentiviral system provided stable and effective gene delivery to chick embryo-derived cells.
Discussion
The study confirmed that lentiviral vectors carrying the PDX1 gene could be efficiently produced and used to achieve stable gene transfer in chick embryonic cells. This system’s ability to integrate permanently into the host genome ensures consistent gene expression over time without repeated transfection. For functional genes like PDX1, this stability is crucial for maintaining insulin-related pathways and pancreatic cell differentiation.
Chick embryos serve as an advantageous model because their cells are easily accessible, grow rapidly, and respond well to viral vectors. Such characteristics make them ideal for investigating genetic regulation during early development.
Evaluation of viral titers using fluorescence microscopy and flow cytometry provided reliable data confirming efficient vector production. The integration of new tools such as CRISPR/Cas9 can further enhance lentiviral design precision, allowing targeted modification of specific genes. Combining these technologies may open promising avenues for studying metabolic disorders and for gene-based therapies.
Conclusion
Recombinant lentiviral vectors carrying the PDX1 gene were successfully generated and used to transduce chick embryonic fibroblast and germ cells. The system exhibited high production efficiency, stable gene expression, and suitability for in vitro studies. These findings demonstrate that lentiviral vectors represent a powerful and versatile platform for gene transfer and experimental modeling in avian systems. Moreover, coupling lentiviral vectors with genome editing technologies could expand future applications in regenerative medicine and genetic engineering.

Abstract Introduction: Concern over fossil energy costs and environmental deterioration, along with energy security, has created a strong motivation for research and development of routes to provide sustainable, renewable fuels. In recent years, the use of biomass to produce highly valued chemicals has attracted widespread attention. lignocellulosic biomass, as a promising renewable resource for biofuel production, has distinct advantages in terms of economic and environmental aspects. The conversion of renewable raw materials to hydrocarbon fuels is an attractive alternative to fossil fuels from an economic and environmental point of view. The production process of lignocellulosic biomass mainly consists of biomass accumulation, biomass decomposition, simple sugars, and conversion of sugars to biofuel. One of the crucial steps in the economic success of lignocellulosic biofuels depends on the inhibition of competitive metabolism in microorganisms to achieve high productivity. To date, a growing focus is on the use of S. cerevisiae and E. coli as cell lines. These two cellular factories have the benefits that are well known.
Aim: The organic compound D-1,2,4-Butanetriol is a valuable chemical with wide-ranging applications in various fields. The chemical synthesis routes for BT have many drawbacks. By genetically modifying microorganisms, the metabolic pathway for producing many substances, including BT, can be engineered. The organic compound D-1,2,4-butanediol (BT) is an important intermediate chemical widely used in fields such as pharmaceuticals, paper, polymer materials, and military applications. When D-xylose sugar is provided to the bacterium, it is first converted to an intermediate compound called xylonolactone. This compound itself slowly converts into xylonate through a non-enzymatic reaction. To produce xylonate, the engineered bacteria have received xylose and initially, by a dehydrogenase reaction by the xylose dehydrogenase enzyme, that converts it into an intermediate substance: xylonolactone. The xylonolactone is converted slowly and in a nonenzymatic reaction to xylonate. Xylonate is a five-carbon organic acid. Over the past few years, xylonate has been increasingly being considered as an important chemical due to its potential as an important chemical component. Xylonate has many applications that can be used in the food, chemical, and pharmaceutical industries. Specifically, xylonate can act as a precursor for the synthesis of D-1,2,4-Butanetriol and a decrease in concrete water. E.coli due to fast growth in cheap culture medium, having two enzymes for the bto synthesis and product production in lower 24 hours of fermentation was chosen as the target strain of genetic engineering and metabolism. This study aimed to clone and express xylose dehydrogenase from Caulobacter vibrioides in E.coli. Materials and Methods: At first, to access the bacterial gene sequence, the genome of the target bacterium was extracted. Then, to create a strain expressing the enzymes xylose dehydrogenase and xylonolactonase, the genes for these proteins were amplified from Caulobacter vibrioides CB1 and transferred to E. coli. For this purpose, the target genes were amplified using specifically designed primers via the Polymerase Chain Reaction (PCR) method and initially cloned into a pTZ57cloning vector and then subcloned into pET 26b expression vector. At the final step, the expression of the enzyme was assessed by SDS-PAGE, and the other confirmation was the reduction of NAD+ to NADH, which was used as an activity indicator of the enzyme, as investigated by a change in NADH absorbance at 340 nm. Results: Confirmatory tests were performed to ensure the presence of the gene in the vectors (using restriction enzymes and colony PCR for gene amplification). The expression and activity of the enzyme were analysed. The recombinant protein's presence was confirmed by SDS-PAGE for the xylose dehydrogenase gene, with a molecular weight of 52.2 kDa. The estimated recombinant protein expression levels were approximately 25%. Conclusion: The objective of this research was solely to establish the metabolic pathway for xylonate production in E. coli by surface expression of enzymes in this pathway (xylose dehydrogenase). The results obtained in this study confirm that half of the pathway is active at the cell surface, but further experiments are required to determine the precise production levels and complete the pathway. This study aimed to create a metabolic pathway for producing xylonate in E.coli.

Abstract Introduction: Pancreatic cancer is the fourth leading cause of cancer-related deaths worldwide. 5-Fluorouracil is one of the commonly used chemotherapeutic drugs. The plant Artemisia absinthium has attracted attention as a potential herbal anticancer agent. This study investigated the effect of the methanolic extract of this plant on the expression of miR-34a and the apoptotic genes BAX, Caspase-3, and Caspase-9 in AsPC-1 pancreatic cancer cells.
Aims: This study aimed to evaluate the cytotoxic and pro-apoptotic effects of Artemisia absinthium methanol extract on pancreatic cancer cells. The research specifically investigated the molecular mechanism by analyzing expression changes in the tumor suppressor miR-34a and key apoptotic genes BAX, Caspase-3, and Caspase-9 to elucidate the extract's anti-cancer mode of action.
Materials and methods: To evaluate the cytotoxic effects of the plant extract on cancer cells, an MTT assay was performed to determine the viability and survival rate of the cells following treatment with various concentrations of the extract, the chemotherapeutic drug fluorouracil (5-FU), and the combined treatment of the extract and the drug. This assay measures cellular metabolic activity and allows quantification of live and dead cells after exposure to different treatments. Based on the obtained results, the IC₅₀ value for each treatment was calculated, representing the concentration at which 50% of the cells were inhibited or killed.
After determining the IC₅₀ value, cells were treated with concentrations equal to, lower, and higher than the IC₅₀ to further investigate the cytotoxic effects and the mode of cell death induced by the treatments. To distinguish between apoptotic and necrotic cell death, the Annexin V-FITC/PI assay was employed. This assay detects phosphatidylserine externalization on the cell membrane and enables differentiation between live, early apoptotic, late apoptotic, and necrotic cells.

In addition to the morphological and physiological assessments, molecular analyses were conducted to examine the expression levels of key apoptosis-related genes, including Caspase-3, Caspase-9, and BAX, as well as the regulatory microRNA miR-34a. Gene expression analysis was performed using Real-time PCR (qPCR).

Results: The results of the MTT assay demonstrated that the proliferation of AsPC-1 pancreatic cancer cells was inhibited by treatment with the extract of Artemisia absinthium and the chemotherapeutic drug fluorouracil (5-FU) in a concentration-dependent manner. As the concentration of each treatment increased, cell viability significantly decreased, indicating a marked cytotoxic effect of both the plant extract and the drug. Moreover, a possible synergistic effect between the extract and fluorouracil in suppressing. To determine the mode of cell death induced by these treatments, the Annexin V-FITC/PI assay was performed. The results revealed that a considerable proportion of treated cells underwent programmed cell death (apoptosis), while the percentage of necrotic cells remained relatively low. These findings suggest that the observed reduction in cell viability is mainly mediated through the activation of apoptotic pathways rather than necrosis. Furthermore, Real-time PCR analysis showed a significant upregulation in the expression of the regulatory microRNA miR-34a and the apoptosis-related genes Caspase-3, Caspase-9, and BAX in the treated groups compared to the control group.
Discussion: The obtained results suggest that Artemisia absinthium extract exerts its cytotoxic effect primarily through the induction of apoptosis rather than necrosis in AsPC-1 pancreatic cancer cells. The observed upregulation of miR-34a, Caspase-3, Caspase-9, and BAX implies activation of intrinsic apoptotic pathways. These findings are consistent with previous studies reporting pro-apoptotic properties of A. absinthium and other Artemisia species. Therefore, the extract may enhance the therapeutic response of pancreatic cancer cells when combined with conventional chemotherapeutic agents such as fluorouracil.
Conclusion: In conclusion, Artemisia absinthium extract demonstrated strong antiproliferative and apoptosis-inducing effects on AsPC-1 cancer cells in a dose-dependent manner. Its combination with fluorouracil produced a synergistic cytotoxic impact, significantly enhancing cell death through apoptotic signaling. The molecular findings support the potential of this extract as a complementary therapeutic agent. Further studies are recommended to explore its mechanisms and evaluate its efficacy in in vivo models of pancreatic cancer.

Abstract Aim: The aim of this study was to investigate different concentrations of iron oxide nanoparticles in and benzylaminopurine (BAP) bell pepper anther culture on the traits of callus formation, embryogenesis, regeneration and rooting.
Material and methods: The experiment was conducted as a factorial experiment in a completely randomized design under in vitro culture conditions. Flower buds of appropriate size (equal sepal to petal ratio or slightly longer petal) were collected from the greenhouse and acetocarmine solution was used to determine the growth and development stage of microspores. The results showed that the most suitable stage for embryogenesis induction was the late mononuclear stage and the early binuclear stage. In order to sterilize the flower buds, 70% ethanol for 30 seconds and 5% sodium hypochlorite for 20 minutes were used and after each stage they were washed three times with sterile distilled water. Then, the anthers were separated from the flower bud and placed in C medium containing 2 mg/L naphthalene acetic acid (NAA), different concentrations of BAP (0, 0.1, 0.5, and 1 mg/L) and different concentrations of iron oxide nanoparticles (0, 1, 10, and 20 mg/L). After that, the explant cultured in C medium were kept at 35°C in the dark for 8 days in order to apply heat treatment. Then, they were transferred to 25°C in the light for 4 days. After this period, in order to induce embryogenesis, the explants were transferred from C medium to R medium and were subcultured every three weeks until embryos emerged. For further growth and root development, the embryos were transferred to V medium. Results: The results of the analysis of variance of the data showed that using different concentrations of iron oxide nanoparticles had a significant effect on the percentage of embryogenesis, regeneration and rooting of the plant, but did not have a significant effect on the percentage of callus formation. The results of comparing the average data showed that among the different concentrations of iron oxide nanoparticles, the 1 mg/L treatment had the highest percentage of embryogenesis (11.11). Also, the results of comparing the average effect of the nanoparticles on the percentage of regeneration showed that the 1 mg/L treatment had the highest percentage of regeneration (16.66). The results of the interaction effects showed that among the different concentrations of iron oxide nanoparticles and BAP, the highest percentage of embryogenesis was observed in the treatment of 20 mg/L iron oxide nanoparticles and 0 mg/L BAP. Also, the treatment of 20 mg/L iron oxide nanoparticles and 0 mg/L BAP had the highest percentage of regeneration (33.33%). After sufficient growth and root formation, the obtained plants were removed from the glass culture containers and transferred to pots containing sterilized culture medium and watered for adaptation. The tops of the pots were covered with plastic cups, and after three days, the cups were pierced and the plastic was gradually removed from the plant for further adaptation. Ploidy levels were determined by chromosome counting by staining the root tip cells. The results showed that out of the 23 obtained plants, 21 were diploid and had 2n=2x=24 chromosomes and 2 were haploid and had n=x=12 chromosomes. Conclusion: In bell pepper anther culture, different concentrations of iron oxide nanoparticles along with plant growth regulators at different concentrations showed a great effect on embryogenesis, regeneration, and rooting.

Abstract During the past 30 years, researchers have made a remarkable progress in identifying the biological (bacteria, viruses), biochemical (chemical compounds) and biophysical (ionizing radiation) cause of human cancer. The term ˝Cancer˝ refers to 277 forms of cancer diseases.
Scientists have determined the process of cancer formation from a consequence of accumulating multiple mutations in human genome. These genetic disruptions would eventually change the normal process of cellular proliferations and differentiation.
The genetic alterations are frequently indicative of poor prognosis for most human cancers.
Both nonhereditary and hereditary cancers are caused by genetic disorders that change the cellular growth control system.
 Genes associated with human cancer formation include four classes of genes: 1. Oncogenes (which increasing their activities end to uncontrollable growth of cells), 2. Tumor suppressor genes, 3. DNA repairing genes, 4. Apoptotic genes.
Over activated oncogenes which cause cellular proliferation. In contrast, inactivated tumor suppressor genes lose their inhibitory effect which is crucial to prevent inappropriate growth. DNA repairing proteins fix the damage and apoptotic proteins cause the pre-cancer cell to commit suicide.
We have over millions of genes in each somatic cell of our body. After sequencing all human genome in 2003, we noticed that Only 23,500 genes are active which encode over 400,000 proteins needed for physiological functions.
99.9% of genome is identical in all humans worldwide. Only 0.1% of the whole genome differ which cause the genetic variations.
Up to 93% of all human cancers are non-hereditary and the remaining 7% are hereditary. A wealth of information has been indicated by the potential use of bioinformatics and molecular techniques for cancer screening, prognosis and monitoring of the efficacy of anticancer therapies. In recent years, molecular genetics have greatly increased our understanding of the basic mechanisms in cancer development. The essential outcome of these molecular studies is that the cancer can be considered as the genetic disease.
 

Abstract Aim: Pentoxifylline (PTX) is a methylxanthine derivative medicine used to improve motility of human spermatozoa in-vitro. It is commonly used in treatment of male-factor infertility, including asthenozoospermia. This study aimed to evaluate the effect of PTX on human sperm parameters and DNA integrity from asthenozoospermic problem.
Material and methods: A total of 38 infertile men with asthenozoospermia were allocated in this experimental study.  Specimens were randomly divided into experimental group treated with 3.6 mM PTX, and control group. All samples were incubated at 37˚ C for 45 min. Semen parameters and sperm DNA fragmentation were measured using sperm chromatin dispersion (SCD) test.
Results: PTX improved sperm motility, significantly, compared to the control (85.76±5.96 Vs 79.44±9.37, respectively, p < .01). There was also a significant decrease in sperm viability in the PTX- treated group in comparison to controls (87.7±8.3 Vs 83.5±9, respectively, p < .01). In addition, sperm DNA fragmentation was higher in PTX-treated group compared to control (23.36±10.25 and 18.5±8.74, respectively, p < .0001).
Conclusion: PTX might have some negative impact(s) on sperm DNA quality, although it has improved the sperm motility. Further studies are needed to elucidate the safety of PTX treatments in ART clinics.

Abstract Aim: The aim of this study was to investigate the effect of vitamin E on sperm parameters in adult rat treated with para-nonylphenol.
Material and methods: Adult male rats were divided into four groups: Control, para-nonylphenol, vitamin E and para-nonylphenl+vitamin E. Oral treatments were performed till 56 days. At the end of treatments, body and left testis weight were recorded and left caudal epididymis was cut in a medium. Released spermatozoa were used to analyze sperm parameters such as sperm number, viability, morphology and motility. Sperm chromatin quality was assessed by nuclear staining using acridine orange and aniline blue.
Results: Body and testis weight as well as normal sperm morphology showed no significant change in four groups. A significant decrease in the number, viability and motility of the sperm was found in rats treated by para-nonylphenol compared to the control. This decrease was significantly compensated by vitamin E in para-nonylphenol+vitamin E group compared to para-nonylphenol group. The application of vitamin E alone could significantly increase sperm viability and motility as comared with the control. Para-nonylphenol had no effect on sperm DNA integrity and histon-protamine replacement compared to the control.
Conclusion: Vitamin E, as a potent antioxidant, could protect the adverse effect of para-nonylphenol on certain sperm parameters in adult rats.
 

Abstract Plants produce a big group of secondary metabolites which are used as medicinal compounds. According to recent estimates, global market value of herbal medicines, including medicinal plants and their products, significantly has been increasing. Considering to the fact that most of the world market for medicinal plants, production and supply of secondary metabolites derived from these plants are concerned and the plant secondary metabolites are of high economic value. Chemical synthesis of these metabolites is an expensive process. So production of metabolites by different biotechnological methods such as cell culture is a useful alternative. Molecular recognition and elicitor-plant receptors interaction is a complex process requiring for signal transduction. Biotic elicitors induce secondary metabolites and hypersensitive responses by activation of defense mechanisms. Manipulation of cell culture media by elicitors is an important strategy for inducing secondary metabolism and production of valuable metabolites. Molecular recognition and elicitor-plant receptors interaction is a complex process requiring for signal transduction. Following perception of elicitor signals, rapid defense responses can be organized as follows: increase of ionic currents across the plasma membrane, reactive oxygen species (ROS) production, activation of defense gene expression, structural changes in the cell wall and phytoalexin production. In this study, different aspects of increasing the production of secondary metabolites in cell culture of plants by biotic elicitors is investigated.  

Abstract Cryopreservation is the most effective method for long-term maintenance of sperm. There are several procedures for freeze-thawing of semen and each may impose damage on sperm function, viability and finally decreases semen quality and fertility potential. In addition to decreased percentage of sperm viability and motility after freeze-thawing, percentage of DNA damage is also increases due to high level of oxidative stress. To minimize these damages, we need to increase our insights regarding different cryopreservation procedures, cryoprotectant and antioxidant supplements, which can protect sperm membrane during cryopreservation. Therefore, by using these experiments, we can improve the efficiency of these procedures. In this review, we discuss about principles of cryopreservation, types of freeze-thawing methods, advantages and disadvantages of each of these methods, effects of freezing on sperm parameters and clinical outcomes, and finally role of antioxidants in preservation of sperm integrity during freeze-thawing. For this review, all relevant information was collected via databases such as PubMed and Google Scholar during the period of 1966-2017.