Cell and Tissue Journal

Research - Scientific -

Examining the expression changes of BAX, Caspase 9, Caspase-3, miR-34a in pancreatic cancer cells of PANC-1 cell line treated with silver nanoparticles synthesized by wormwood plant (Artemisia absinthium)

Pages 104-120

https://doi.org/10.66224/JCT.17.2.104

Seyyed Morteza Mortazavi, Khadije Nejad Shahrokhabadi, javad baharara, Maryam Lotfi

Abstract Introduction: Concerns about fossil energy costs, environmental deterioration, and energy security has created strong motivation for the research and development of routes to provide sustainable and 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 benefits. The conversion of renewable raw materials to hydrocarbon fuels is an attractive alternative to fossil fuels from economic and environmental perspectives. 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 for the economic success of lignocellulosic biofuels depends on the inhibition of competitive metabolism in microorganisms to achieve high productivity. To date, there has been a growing focus on the use of S. cerevisiae and E. coli as cell lines. These two cellular factories have well known advantages. They are genetically transmissible and several tools are available for genetic manipulation. In order to produce xylonate, the engineered xylose is first converted by a dehydrogenase into the intermediate xylonolactone, which is then slowly converted to xylonate in a nonenzymatic reaction.
Aim: The organic compound D-1,2,4-Butanetriol (BT) is a valuable chemical with wide-ranging applications in various fields such as pharmaceuticals, paper, polymer materials, and military applications. However, 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. When D-xylose is supplied to the bacterium, it is first converted into an intermediate compound called xylonolactone. This compound slowly converts into xylonate through a non-enzymatic reaction. To produce xylonate, the engineered bacteria receive xylose, which is initially converted by a dehydrogenase reaction catalysed by the xylose dehydrogenase enzyme into an intermediate compound, xylonolactone. Xylonolactone is slowly converted to xylonate in a nonenzymatic reaction. Xylonate is a five-carbon organic acid. Over the past few years, xylonate has increasingly been considered as an important chemical due to its potential as an important chemical component. Xylonate has many applications in the food, chemical, and pharmaceutical industries. Specifically, xylonate can act as a precursor for the synthesis of D-1,2,4-Butanetriol and as a concrete water reducing agent. E. coli was chosen as the target strain for genetic and metabolic engineering due to its fast growth in inexpensive culture media, the presence of two enzymes for BT synthesis, and product formation in less than 24 hours of fermentation. 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 into 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 analyzed. 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 expression level of the recombinant protein was 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.

Research - Scientific -

Comparison of the Effects of Papaverine and Tamoxifen on the Expression of CDK4, miR-146a, and miR-22 in DU145 Prostate Cancer Cell line

Pages 161-173

https://doi.org/10.66224/JCT.17.2.161

Hamid Reza Momeni, Tahereh Etemadi, Arkan Al Masoodi, Zahra Azizi

Abstract Introduction: Prostate cancer is one of the most prevalent malignancies among men worldwide and represents a major cause of cancer-related morbidity and mortality. Despite advances in screening and therapeutic approaches, resistance to standard treatments, including androgen‑deprivation therapy, remains a major clinical challenge in prostate cancer. As a result, identifying novel compounds that can suppress proliferation or modulate key molecular regulators of the cell cycle is of considerable interest. Papaverine is an isoquinoline alkaloid derived from the opium poppy, widely known for its long‑standing clinical use as a smooth‑muscle relaxant and vasodilator. In recent years, it has gained attention for its potential anticancer effects, including its influence on mitochondrial function, cellular energy metabolism, and the modulation of signaling pathways involved in cell cycle regulation. However, the specific effects of papaverine on prostate cancer cells have not yet been clearly elucidated. Tamoxifen, a selective estrogen receptor modulator widely used in breast cancer therapy, has also demonstrated off-target antiproliferative effects in various tumor models.
Aims: The present study aimed to evaluate the effects of papaverine and tamoxifen on the viability and molecular regulatory profile of the DU145 prostate cancer cell line. Specifically, the study investigated the cytotoxic potential of these compounds at different concentrations and incubation times, and their ability to alter the expression of the CDK4 gene, a key mediator of G1-to-S phase progression, along with two microRNAs, miR-146a and miR-22, known to influence cell cycle control and tumorigenic pathways. By integrating cellular and molecular findings, the study sought to clarify whether these compounds could serve as potential modulators of prostate cancer cell growth.
Materials and Methods: The DU145 human prostate cancer cell line was cultured under standard conditions and treated with different concentrations of papaverine and tamoxifen. Cell viability was assessed for 24, 48, and 72 hours using the MTT (3- (4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide) assay. Dose–response curves were generated to calculate IC50 values for each compound at the specified time points. To investigate molecular changes, total RNA was extracted from treated and control cells, and the expression levels of the CDK4 gene, miR-146a, and miR-22 were quantified by Real-Time PCR with appropriate internal controls. Relative expression changes were determined using the 2^-ΔΔCt method. The data were analyzed using SPSS software. Results were presented as Mean ± SD, and a significance level of p ≤ 0.05 was considered statistically significant.
Results: Treatment of the DU145 prostate cancer cell line with various concentrations of papaverine and tamoxifen resulted in a significant reduction in cell viability in a concentration- and time-dependent manner, as measured by the MTT assay for 24, 48, and 72 hours. IC50 values were successfully determined for both compounds across the different time points.
Molecular analysis using Real-Time PCR showed that treatment with either papaverine or tamoxifen led to a marked downregulation of CDK4 expression compared with untreated controls. In contrast, both compounds induced a significant up‑regulation of miR‑146a and miR‑22 levels.
Discussion: The observed decrease in viability suggests that papaverine and tamoxifen exert notable antiproliferative effects on DU145 prostate cancer cells. The downregulation of the CDK4 gene provides a mechanistic link to cell cycle arrest, as CDK4 is a central regulator of G1‑to‑S phase progression. Such suppression is in line with reduced cellular proliferation and supports the therapeutic potential of these compounds.
The increased expression of miR‑146a and miR‑22 further highlights the involvement of tumor suppressive microRNAs in mediating these effects. Both miRNAs have been implicated in regulating pathways associated with proliferation, apoptosis, and oncogenic signaling. Their upregulation may contribute to the inhibition of cell growth through negative modulation of critical oncogenic targets.
These results align with prior evidence demonstrating tamoxifen’s antiproliferative effects, but also importantly emphasize that the molecular and cellular impacts of papaverine in prostate cancer remain poorly understood. The current findings provide initial insight into papaverine’s interaction with cell cycle regulatory genes and microRNAs, suggesting a potential role in prostate cancer growth inhibition. Considering the adverse effect profile associated with tamoxifen, papaverine may therefore represent a more favorable candidate for further investigation, particularly as a potential alternative with a potentially safer therapeutic window.
Conclusion: In conclusion, both papaverine and tamoxifen effectively reduced DU145 cell viability and modulated the expression of the CDK4 gene, miR‑146a, and miR‑22, indicating their capability to interfere with cell cycle regulatory mechanisms. While their antiproliferative effects were broadly comparable, the established side effects of tamoxifen suggest limitations in its clinical applicability. Given that papaverine produced similar molecular outcomes without the same known adverse profile, it may be considered a potentially more suitable candidate for further preclinical evaluation in prostate cancer research.

Research - Scientific -

Engineering of CNC-based hydrogel nanoparticles with controlled concentrations to enhance the selectivity and efficacy of cisplatin in AGS cells

Pages 174-194

https://doi.org/10.61882/jct.2026.2088522.2135

Payam Madazadeh, hashem yaghoubi

Abstract Introduction: Cancer is recognized as one of the leading causes of mortality worldwide and remains a major global health challenge. Among the various types of cancer, gastric cancer is considered one of the most aggressive and prevalent malignancies due to its high incidence rate, late diagnosis, and poor prognosis. The AGS cell line is widely used as a reliable in vitro model for studying human gastric adenocarcinoma because of its adhesive growth pattern, relatively rapid proliferation, and preservation of the invasive characteristics of primary tumor cells. Conventional therapeutic approaches for gastric cancer, including surgery, radiotherapy, and chemotherapy, have shown partial clinical success; however, these methods are frequently associated with serious limitations such as systemic toxicity, damage to healthy tissues, severe side effects, and the development of drug resistance. Consequently, the development of advanced drug delivery systems has attracted considerable attention as a promising strategy to improve therapeutic efficacy while minimizing adverse effects. In this regard, nanotechnology has emerged as a powerful platform for the design of smart nanocarriers capable of enhancing drug stability, improving bioavailability, and enabling controlled and targeted drug release. Cisplatin is one of the most important chemotherapeutic agents used in the treatment of gastric cancer and exerts its anticancer activity primarily through binding to DNA, disrupting replication and transcription processes, and ultimately inducing apoptosis. Nevertheless, its clinical application is limited by severe toxicity and acquired drug resistance. Cellulose nanocrystals have gained significant attention in drug delivery systems owing to their biodegradability, high mechanical strength, structural stability, and ability to provide sustained and controlled drug release. In addition, bovine serum albumin is considered an ideal biological carrier because of its excellent biocompatibility, low toxicity, biodegradability, and natural tendency to accumulate in tumor tissues. Polyvinyl alcohol also plays an important role as a biocompatible polymeric matrix in the fabrication of stable nanocarriers. Furthermore, hyaluronic acid serves as a targeting ligand through its specific interaction with CD44 receptors, which are overexpressed on the surface of many cancer cells, including gastric cancer cells. This targeted interaction facilitates the selective uptake of nanoparticles by tumor cells and significantly enhances the efficiency of targeted drug delivery systems.
Aims: The aim of this study was to design and evaluate a smart nanocarrier system based on cellulose nanocrystals, polyvinyl alcohol, bovine serum albumin, and hyaluronic acid for the targeted delivery of cisplatin to gastric cancer cells. This study sought to improve drug stability, controlled release behavior, selective cellular uptake, and reduced toxicity toward normal cells by utilizing the biocompatible and targeting properties of these components. In addition, the cytotoxicity and anticancer selectivity of the synthesized nanoparticles were investigated in the AGS gastric cancer cell line to evaluate the potential efficiency of this nanocarrier system in enhancing the therapeutic performance of gastric cancer treatment.
Materials and methods: In this study, cellulose nanocrystals were extracted from raw cellulose through controlled sulfuric acid hydrolysis using 64% sulfuric acid, followed by ultrasonication, centrifugation, and dialysis to obtain both stable suspensions and freeze-dried powders. Cisplatin was subsequently loaded onto cellulose nanocrystals at different concentrations, and the drug-loading efficiency was determined spectrophotometrically at 370 nm. Multifunctional BSA (Bovine Serum Albumin)-PVA (Polyvinyl Alcohol)-CNC (Cellulose Nanocrystals)-CDP (Cisplatin) nanoparticles coated with PEG-hyaluronic acid were then synthesized by initially coating the drug-loaded cellulose nanocrystals with bovine serum albumin, followed by the formation of a stable hydrogel network using polyvinyl alcohol. To improve targeting ability and biological stability, the nanoparticle surface was further functionalized with NH2-PEG-HA in the presence of EDC/NHS as a coupling agent, and the final suspension was purified by dialysis before further analyses. The physicochemical properties of the synthesized nanoparticles, including crystalline structure, thermal stability, particle size, and zeta potential, were characterized using X-ray Diffraction (XRD), Thermogravimetric Analysis (TGA), and Dynamic Light Scattering (DLS) analyses, respectively. In addition, the release kinetics of cisplatin from the nanocapsules were investigated under different pH conditions simulating normal and tumor microenvironments, and the obtained data were analyzed using the Korsmeyer–Peppas kinetic model. To evaluate biocompatibility and selective cytotoxicity, AGS gastric cancer cells and normal GES-1 gastric epithelial cells were treated with the synthesized nanocapsules and free cisplatin, followed by MTT assay for IC₅₀ determination. Furthermore, the selectivity index (SI) was calculated to assess preferential toxicity toward cancer cells. Finally, apoptosis induction was evaluated using Annexin V/PI staining and flow cytometry. All experiments were performed in triplicate, and the data were statistically analyzed using one-way ANOVA followed by Duncan’s multiple range test in SPSS22 software.
Results: The physicochemical characterization results demonstrated that all synthesized nanoparticles and nanocapsules possessed a predominantly amorphous structure. The XRD patterns exhibited broad diffraction peaks within the 15–30° range, while the absence of sharp crystalline peaks confirmed the successful incorporation of cellulose nanocrystal–cisplatin complexes into the BSA-PVA matrix and the formation of a homogeneous noncrystalline system. TGA analysis further revealed that increasing the concentration of cellulose nanocrystals from 50 to 200 mg significantly enhanced the thermal stability of CNC-CDP nanoparticles, as higher concentrations remained thermally stable up to nearly 300°C. Moreover, the BPC_{(100 mg)}CP-HA nanocapsules showed lower thermal degradation and greater structural stability compared with drug-free nanocapsules, indicating the reinforcing effect of CNC-CDP within the nanocapsule network. DLS analysis demonstrated that increasing the cellulose nanocrystal concentration increased particle size from 322 to 363 nm, while the zeta potential decreased from +13.05 to +2.14 mV. Surface coating with BSA, PVA, PEG, and hyaluronic acid further increased the particle size to 476 nm and elevated the PDI value, confirming the successful formation of multilayered nanocapsules. Drug release studies indicated that cisplatin release was significantly higher under acidic conditions (pH=5.8) than under physiological conditions (pH=7.4), and the release rate increased over time. In contrast, increasing the cellulose nanocrystal concentration reduced the drug release rate, suggesting a more compact and stable nanostructure. MTT assay results showed that drug-free nanocapsules exhibited negligible cytotoxicity against both AGS and GES-1 cells, whereas cisplatin-loaded nanocapsules significantly reduced the viability of cancer cells. Among all formulations, BPC_{(50 mg)}CP-HA exhibited the strongest anticancer activity, with an IC₅₀ value of 224.75 µg.mL^-1 against AGS cells and the highest selectivity index (SI=1.75), indicating superior selective toxicity toward cancer cells. Furthermore, flow cytometry analysis demonstrated that this formulation induced the highest percentages of early and late apoptotic cells, while free cisplatin caused the greatest level of necrotic cell death.
Discussion: In recent years, smart nanocarrier systems have emerged as promising strategies for targeted cancer therapy due to their ability to improve drug stability, provide controlled release, and reduce systemic side effects. In the present study, multifunctional nanocapsules composed of cellulose nanocrystals, BSA, PVA, PEG, and hyaluronic acid were designed for the targeted delivery of cisplatin to AGS gastric cancer cells. The obtained results demonstrated that drug-free nanocapsules exhibited negligible cytotoxicity against both AGS and GES-1 cells, indicating the favorable biocompatibility of the synthesized nanocarrier system. This biocompatibility may be attributed to the intrinsic properties of the nanocarrier components, which are unlikely to induce oxidative stress, membrane damage, or mitochondrial dysfunction in healthy cells. Furthermore, PEGylation and hyaluronic acid-mediated targeting reduced nonspecific nanoparticle accumulation in normal cells while improving selective uptake by cancer cells through CD44 receptor interactions. Physicochemical characterization revealed that increasing the concentration of cellulose nanocrystals resulted in larger particle sizes and lower zeta potential values, likely due to the formation of denser polymeric networks and stronger hydrogen-bonding interactions among the nanocarrier components. Nevertheless, the BPC_{(100 mg)}CP-HA formulation exhibited an appropriate particle size distribution and acceptable uniformity for drug delivery applications. Drug release studies further demonstrated that cisplatin release was significantly higher under acidic conditions than at physiological pH, while increasing cellulose nanocrystal concentration reduced the release rate. This behavior was associated with the formation of a more compact polymeric network that restricted drug diffusion and enabled sustained and controlled release. In addition, flow cytometry analysis revealed that the BPC_{(50 mg)}CP-HA nanocapsules induced the highest levels of apoptosis in AGS cells, whereas free cisplatin predominantly caused necrotic cell death. These findings suggest that encapsulation of cisplatin within the designed nanocarrier system not only enhances anticancer selectivity but also promotes apoptosis-mediated cell death, thereby potentially improving the therapeutic efficacy of cisplatin in gastric cancer treatment.
Conclusion: This study demonstrates that smart nanocapsules BPCCP-HA based on cellulose nanocrystals and a BSA-PVA hydrogel provide an effective strategy to overcome the therapeutic limitations of cisplatin. This system enhances physicochemical properties, structural stability, and pH-responsive drug release, thereby improving targeted delivery of the drug to AGS cancer cells. The designed formulation shows good biocompatibility and low toxicity toward healthy cells.

Review Articles -

Studying the molecular and physiological mechanisms of glycine betaine accumulation in order to improve tolerance to abiotic stresses in plants

Articles in Press, Accepted Manuscript, Available Online from 13 May 2026

https://doi.org/10.61882/jct.2026.2081953.2123

Lavin Babaei, Reza Darvishzadeh

Abstract Introduction: Global climate change has significantly increased the frequency and intensity of abiotic stresses, thereby limiting plant growth, development, and overall yield by damaging physiological systems. In response to environmental stressors, plants have evolved various adaptive mechanisms, including the accumulation of compatible solutes such as glycine betaine (GB), which plays a pivotal role in protecting cellular functions under adverse conditions. Also known simply as betaine, this compound is a methylated glycine derivative recognized across plant species for its ability to mitigate the deleterious effects of stressful environments. Its zwitterionic structure; comprising a positively charged trimethylammonium group and a negatively charged carboxyl group, confers high solubility and chemical stability to the molecule.
Owing to its excellent biocompatibility, favorable carbon-to-nitrogen ratio, and high-concentration accumulation, glycine betaine can enhance plant tolerance against a wide spectrum of abiotic stresses. Specifically, GB contributes to photosynthetic recovery and the alleviation of oxidative stress by reducing the accumulation and facilitating the detoxification of reactive oxygen species (ROS). Furthermore, it plays a crucial role in stabilizing membranes and macromolecules, while protecting key components of the photosynthetic apparatus, such as the Rubisco enzyme, Photosystem II (PSII), quaternary enzymes, and complex protein structures. Notably, glycine betaine can accumulate at high concentrations within plant cells without interfering with normal metabolic processes, thereby significantly increasing resilience to various osmotic stresses, extreme temperatures (heat and cold), and oxidative damage. The biosynthesis of glycine betaine occurs through distinct metabolic pathways, including the choline oxidation pathway (prevalent in plants and mammals), the direct glycine methylation pathway (specific to certain bacteria and halophytes), the choline dehydrogenase pathway, and the serine metabolism pathway. Such diversity underscores the vital importance of this osmolyte in mediating responses to environmental stresses. Recently, biotechnological interventions, such as Agrobacterium-mediated transformation, have successfully enhanced stress tolerance in susceptible species by overexpressing key genes, most notably codA, BADH, GSMT, and SDMT. Given the functional diversity of genes involved in the glycine betaine biosynthetic pathway, extensive efforts have been made to develop transgenic plants capable of effective accumulation of this metabolite; however, serious challenges such as unstable and weak transgene expression remain as key obstacles in this path. Factors including promoter type, genomic integration site, and epigenetic factors can influence the final performance. Furthermore, the overexpression of enzymes in the glycine betaine biosynthetic pathway may potentially impair growth by disrupting metabolic stability. Therefore, future research should focus on decoding the molecular networks regulating the biosynthesis, signaling, and transport of glycine betaine, particularly its crosstalk with phytohormones, transcription factors, and the identification of stress-inducible promoters. Additionally, optimizing transformation protocols and synchronizing glycine betaine gene expression with the overall plant metabolism are essential.
Aims: This review examines the biosynthesis, physiological functions, and molecular regulation of glycine betaine (GB). It highlights genetic engineering techniques to boost GB production, offering sustainable strategies for enhancing crop tolerance to environmental stresses.
Conclusion: In-planta biosynthesis of glycine betaine (GB) offers a more sustainable alternative to exogenous application, aligning closely with the principles of green agriculture. By integrating GB synthesis pathways, genetically engineered crops can autonomously boost metabolite production and bolster stress resilience, thereby eliminating the logistical costs of external treatments. Furthermore, a more profound investigation into these biosynthetic pathways will facilitate the identification and cloning of novel target genes, ultimately maximizing GB accumulation and enhancing environmental tolerance.

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Review Articles -

Bladder cancer treatment with vaccines

Articles in Press, Accepted Manuscript, Available Online from 09 June 2026

https://doi.org/10.61882/jct.2026.2064491.2097

Fatemeh Rezaei, Maedeh Tamimi, mohammad javad dehghan esmatabadi, Ali Asghar Deldar

Abstract Bladder cancer is one of the most common malignancies of the urinary tract and ‎originates from the bladder epithelium. Despite advances in surgery, chemotherapy, ‎radiation, and immunotherapy, bladder cancer remains a major clinical challenge ‎because of its high recurrence rate, risk of progression, and the need for long-term ‎surveillance and repeated interventions. In recent years, vaccine-based immunotherapy ‎has gained increasing attention as a promising strategy for improving antitumor ‎immune responses, reducing relapse, and enhancing therapeutic outcomes across ‎different disease stages.‎
Bacillus Calmette–Guérin (BCG) is the first and most established vaccine used in ‎bladder cancer treatment and remains the standard intravesical adjuvant ‎immunotherapy for non muscle invasive bladder cancer (NMIBC). Its antitumor ‎activity is largely mediated by immunomodulatory effects within the bladder ‎microenvironment, including activation of innate immune cells, induction of ‎pro inflammatory cytokines, and subsequent priming of adaptive immune responses ‎that support cytotoxic T cell–mediated tumor control. However, a substantial ‎proportion of patients experience intolerance, inadequate response, recurrence, or ‎resistance, underscoring the need for next generation vaccine strategies with improved ‎specificity and durability.‎
Accordingly, several emerging vaccine platforms have been investigated. Cell based ‎vaccines, particularly dendritic cell (DC) vaccines, aim to exploit the antigen presenting ‎function of DCs to enhance tumor specific T cell activation using tumor lysates, ‎defined tumor associated antigens, or engineered immunostimulatory constructs. In ‎parallel, monoclonal antibody–based approaches that target tumor associated antigens ‎have been explored as immunotherapeutic tools that may complement vaccine-‎induced immunity. Peptide vaccines provide a more defined strategy by stimulating ‎antigen specific responses against selected epitopes, and their performance may be ‎strengthened through the use of appropriate adjuvants, optimized epitope selection, ‎and improved delivery systems. More recently, mRNA vaccines have attracted ‎considerable interest because they enable rapid and flexible design, can encode one or ‎multiple tumor antigens, and may induce robust cellular immunity. These platforms ‎also offer opportunities for personalization based on patient specific antigenic profiles ‎and may be manufactured in a scalable manner.‎
Combination therapy is an important direction in this field. Pairing vaccine approaches ‎with immune checkpoint inhibitors (e.g., PD 1/PD L1 blockade) may enhance ‎therapeutic efficacy by reversing tumor mediated immune suppression and improving ‎the magnitude and durability of antitumor responses. In addition, integrating vaccines ‎with conventional modalities such as chemotherapy or radiotherapy may further ‎augment immunogenicity through increased antigen release, activation of danger ‎signaling pathways, and immune priming, thereby potentially converting ‎immunologically “cold” tumors into more responsive disease states.‎
This review summarizes major vaccine strategies investigated for bladder cancer, ‎including BCG, cell based vaccines, peptide vaccines, and mRNA vaccines, and ‎discusses their proposed mechanisms, advantages, and current limitations. Despite ‎encouraging progress, key challenges remain, including tumor heterogeneity, antigen ‎loss, immune evasion, optimization of delivery and dosing schedules, manufacturing ‎complexity, and cost. Future research should focus on refining antigen discovery and ‎vaccine design, identifying predictive biomarkers for patient selection and response ‎monitoring, optimizing combination regimens to maximize synergy, and conducting ‎larger, well designed clinical studies to support clinical translation. Overall, vaccine-‎based immunotherapy represents a rapidly evolving and promising avenue that may ‎contribute to more effective, durable, and individualized treatment of bladder cancer ‎for patients across disease stages.‎

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Research - Scientific -

REVIEW SPERM DNA BREAKAGE ININDIVIDUALS WITH OLIGOASTHENOSPERMIA AND ITS EFFECT ON ICSI AND EMBRYO FORMATION IN THE INFERTILITY VENTER ROUYESH KARAJ

Articles in Press, Accepted Manuscript, Available Online from 10 June 2026

https://doi.org/10.61882/jct.2026.2080079.2122

Golriz Lak, farinaz behfarjam

Abstract Introduction: About half of the causes of infertility are related to men. The sperm of infertile men often have different functional and structural defects. Among these defects is sperm DNA damage, which can be caused by DNA fragmentation, improper chromatin packaging, and epigenetic defects. All men have some degree of damage to sperm DNA, but when the percentage of this damage increases significantly, it causes pregnancy disruption, miscarriage, and failure in assisted reproductive methods. Studies have shown that 15% of couples are unable to have children despite trying to conceive and are considered infertile, with half of these cases of infertility being due to male factors. The majority of male infertility is due to abnormal sperm. Therefore, these individuals are candidates for assisted reproductive techniques such as IVF and ICSI and may have tried these treatments repeatedly and failed. One of the most important factors in the success of in vitro fertilization is the health of sperm DNA. Various studies show that the lower the quality of sperm, the more problems sperm DNA health faces.
Aims: Therefore, the purpose of conducting research to identify sperm DNA damage and study its effect on the success rate of assisted reproductive methods, including microinjection, in infertile couples is essential, with the aim of improving sperm quality before starting the treatment cycle and imposing excessive costs on couples.

Materials and methods: The present study was experimental and appropriate laboratory equipment, materials, and solutions were used in the research process. In this experiment, semen samples were collected from 60 infertile couples (30 of which were considered positive control samples) who had referred to the Royesh Infertility Treatment Center in Karaj for ICSI treatment. Oligoasthenospermia samples and positive control samples were selected and separated. Anti-Mullerian hormone (AMH) of women candidates for ICSI, which is present in the patient's serum and measured through a blood test, was examined, and those with AMH equal to 2 and above were selected. Then, sperm washing was performed for intracytoplasmic injection. Sperm injection into the egg is usually done 2-3 hours after ovulation. In the next step, sperm DNA breakage was assessed. Failure assessment was performed immediately after sample receipt. The collected data was analyzed using SPSS software, which provides a summary of the methods and techniques used.
Results: The average embryo quality in the normal sperm group was 93.35%, while this index was only as high as 75.74% in the DNA-damaged sperm group. Also, the average sperm velocity in the normal sperm group was 36.2%, while this index in the DNA-damaged sperm group showed a small value of 5.53%. Also, The average percentage of non-motile sperm, sperm with DNA breakage was 82.17%, and in the normal sperm group, the average was 41.93%. In addition, in the DNA breakage sperm group, the average percentage of sperm with rotational movement was 17%, sperm with slow progressive movement was 5.4%, and sperm with fast progressive movement was 0.1%, while in the normal sperm group, the desired indicators showed averages of 0.87%, 25.13%, and 11.07%, respectively. The results of the study also showed that sperm with DNA breakage has a significant impact on reducing embryo quality. And sperm DNA damage can severely and significantly reduce sperm velocity and count. And finally, sperm DNA breakage has a significant impact on sperm shape.
Discussion: The results obtained indicate that DNA breakage is more common in oligoasthenospermic individuals than in healthy individuals, and this breakage significantly affects embryo quality.
Conclusion: Since in assisted reproductive methods, damage to sperm DNA may cause treatment failure, it is recommended to check the sperm DNA fragmentation rate (SDFA) before choosing the appropriate treatment method. And in many cases, if there is damage to sperm DNA, appropriate treatment can improve sperm quality. SDFA testing is a reliable method for assessing sperm DNA health, which greatly contributes to the clinical diagnosis and treatment of male infertility and is of great value in the success of treatment methods.

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Research - Scientific --

Comparison of Cryopreservation Methods on Sperm Viability in Domestic Animals: A Systematic Review and Meta-Analysis

Articles in Press, Accepted Manuscript, Available Online from 21 July 2026

https://doi.org/10.61882/jct.2026.2089485.2136

Mahdi Khodaei Motlagh, Sobhan Akarim Almdar

Abstract Introduction: Sperm cryopreservation is a critical biotechnology for genetic resource conservation and artificial insemination in domestic animals. However, conventional glycerol-based methods cause severe damage to the plasma membrane, mitochondria, and sperm DNA due to intracellular ice crystal formation, osmotic stress, and increased reactive oxygen species (ROS), ultimately reducing post-thaw viability and fertility. The studies included in this meta-analysis cover multiple economically important species, including cattle, sheep, goats, and particularly poultry (roosters), all of which hold significant value for both economic productivity and biodiversity conservation in Iran. This systematic review and meta-analysis synthesizes evidence from 11 comparative studies to evaluate the efficacy of optimized cryopreservation protocols (e.g., DMA pellets, dry thawing, nano-enhanced extenders, and programmable freezing) versus standard glycerol-based methods on sperm viability and motility.
Aims: The primary objective is to quantify the pooled effect size of optimized cryopreservation methods specifically on post-thaw viability and progressive motility compared to standard glycerol‑based protocols. The secondary objectives are to identify sources of heterogeneity, to assess publication bias, to provide predictions for future studies, and to offer practical, actionable recommendations for implementing these technologies in Iran, particularly for poultry gene banking and the preservation of native breeds.
Materials and Methods: Following PRISMA 2020 guidelines, we systematically searched PubMed, Scopus, Web of Science, Google Scholar, SID, MagIran, and reference lists up to November 2025. Search keywords included: sperm cryopreservation, motility, DMA, nanoparticles, and poultry.
Inclusion criteria were: comparative studies in domestic animals, reporting mean ± SD for post-thaw viability/motility, with a sample size of at least 6 (n ≥ 6). Quality assessment of the included studies was performed using a risk-of-bias tool. Data were analyzed using CMA v4 with a random-effects model. Effect size was calculated as the standardized mean difference (SMD). Heterogeneity was assessed using the statistic and Q-test. Publication bias was evaluated via Funnel Plot and Egger's test.
Results: Eleven studies (comprising 276 sperm samples) were included in the final meta-analysis. Optimized methods significantly improved viability (SMD = 4.514, 95% CI: 3.253–5.776, Z = 7.014, p < 0.001; I² = 86.2%) and motility (SMD = 3.161, 95% CI: 2.378–3.943, Z = 7.919, p < 0.001; I² = 75.7%). The Q-test also revealed significant heterogeneity (p < 0.001), which is entirely expected given the diversity of animal species and cryoprotectant types employed across the primary studies. However, Egger's test indicated no significant publication bias for either outcome (p > 0.05).
Discussion: The pooled analysis showed very large effect sizes favoring optimized methods for both viability and motility. The high heterogeneity was expected and partly explained by species differences and the type of cryoprotectant used. From a biological perspective, DMA offers distinct advantages over glycerol due to its lower molecular weight and faster membrane permeability, which reduces equilibration time and minimizes cytotoxicity. Programmable freezing, by precisely controlling the cooling curve, prevents thermal shock and preserves the structural integrity of the spermatozoa.
Moreover, the incorporation of nanoparticles (e.g., selenium or titanium dioxide) into extenders effectively scavenges ROS, thereby inhibiting lipid peroxidation and optimizing sperm energy metabolism. The dry-thawing method further reduces cold-shock injury by minimizing the time spermatozoa spend in the critical temperature zone (−15 to 0°C), thus preventing recrystallization damage. The findings strongly support the adoption of DMA pellets, dry thawing, and nano‑enhanced extenders in breeding programs and gene banks, especially in Iran.
Conclusion: Optimized protocols substantially outperform standard methods. Given Iran's diverse climatic conditions and the urgent need to protect native poultry breeds from environmental and epidemiological threats, implementing these advanced protocols in national genetic resource centers and the country's gene bank is essential. Furthermore, standardizing rapid dry-thawing protocols across artificial insemination centers nationwide would be a highly effective step toward increasing practical fertility rates, reducing sperm wastage in commercial flocks, and ultimately supporting national food security goals. Therefore, we recommend the widespread adoption of these optimized methods to enhance fertility outcomes and ensure robust genetic preservation in domestic animals.

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Molecular and Cellular Basis of Human Cancer

Volume 2, Issue 4, Winter 2012, Pages 365-376

https://doi.org/10.52547/JCT.2.4.365

N. Parsa

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.

The effect of pentoxifylline on human sperm parameters and DNA integrity– An in-vitro study

Volume 5, Issue 3, Autumn 2014, Pages 281-287

https://doi.org/10.52547/JCT.5.3.281

S Gh, MA Kh, A N, I H, P A, S P

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.

Increasing the Production of Plant Secondary Metabolites Using Biotic Elicitors

Volume 4, Issue 2, Autumn 2013, Pages 119-128

https://doi.org/10.52547/JCT.4.2.119

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.

Protective Effect of Vitamin E on Sperm Parameters in Adult Rat Treated with Para-nonylphenol

Volume 2, Issue 4, Winter 2012, Pages 415-424

https://doi.org/10.52547/JCT.2.4.415

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.

Human sperm cryopreservation update in treatment of infertility: a review study

Volume 8, Issue 4, Spring 2018, Pages 332-353

https://doi.org/10.52547/JCT.8.4.332

B Torki-Boldaji, L Azadi, M Tavalaee, MH Nasr- Esfahani

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.

Evaluation of Curcuma longa L. and bovine ghee effects on experimental stomach ulcers healing in rat

Volume 6, Issue 2, Summer 2015, Pages 123-134

https://doi.org/10.52547/JCT.6.2.123

SE S, Sh E, N M, M F, S N, H R

Abstract Aim: This study was done to evaluate the therapeutic effects of Curcuma longa L. rhizome powder and bovine ghee mixture on experimental stomach ulcers in rat.
Material and methods: Stomach ulcer induction was done in 48 food deprived male rats (250-300 gr) using orally Indomethacin (50 mg/kg suspended in 1% carboxymethyl cellulose. Animals divided randomly into five groups. The normal and untreated (Indomethacin) group just received normal saline. The treated groups received different doses of C. longa and bovine ghee (500 and 750 mg C. longa in 10 ml ghee/per kg) for three successive days. On the third day, the rats were killed and their stomachs were removed for histological studies. Stomach ulcers number and length were measured and also ulcer index was calculated.
Results: Results showed that the mixture of C. longa and bovine ghee is significantly caused decreasing gastric ulcer index, inflammatory cells, blood capillaries densities (p < 0.001) and increasing mucosal layer thickness (p < 0.001) and secreted mucus (p < 0.05) in treated groups in comparison with untreated group.
Conclusion: According to the results, the mixture of C. longa and bovine ghee is significantly accelerated healing of experimental stomach ulcers.

Apoptosis Will be Increased after 2h Incubation of Human Sperm at 37°C

Volume 4, winter, Winter 2014, Pages 415-423

https://doi.org/10.52547/JCT.4.4.415

Abstract Aim: The main goal was to evaluate the impact of different incubation time intervals on human sperm DNA status using terminal deoxyribonucleotidyl transferase–mediated dUTP nick-end labeling (TUNEL) test.
Material and methods: This prospective study involved 21 normozoospermic specimens. After direct swim-up, sperm cells were incubated at 37^°C and DNA damage was evaluated at different time intervals (0, 1, 2 and 3 h). After slide fixation with methanol 100% for 4 minutes and rinse in phosphate-buffered solution (PBS), TUNEL was added to each slide and incubated for 60 minute in dark. Eosin-Nigrosin and Papanicolaou staining protocols were applied in order to assess sperm viability and morphology, respectively.
Results: Sperm viability and normal morphology was improved after sperm processing (100%, 72.3% respectively, p < 0.0001). The rate of DNA damage was significantly higher after 2h compared to 0h (9.19±0.8% Vs 4.9±0.9%, respectively, p=0.008). Also there was significant difference in abnormal sperm DNA between 3h and 1h (10.95±0.7% Vs 7.1±1%, respectively, p=0.020).
Conclusion: Incubation of prepared normozoospermic samples at 37^°C more than 2 h may be associated with sperm DNA fragmentation. Therefore, it seems that incubation of human spermatozoa at 37^°C should be limited up to 2h prior to use in ART clinics.

Number of Volumes	17
Number of Issues	63
Number of Articles	477
Number of Contributors	1,030
Article View	3,067,411
PDF Download	869,863
View Per Article	6430.63
PDF Download Per Article	1823.61

Number of Submissions	1,097
Rejected Submissions	520
Rejection Rate	47
Accepted Submissions	126
Acceptance Rate	11
Time to Accept (Days)	147
Number of Indexing Databases	8
Number of Reviewers	745

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Examining the expression changes of BAX, Caspase 9, Caspase-3, miR-34a in pancreatic cancer cells of PANC-1 cell line treated with silver nanoparticles synthesized by wormwood plant (Artemisia absinthium)

Comparison of the Effects of Papaverine and Tamoxifen on the Expression of CDK4, miR-146a, and miR-22 in DU145 Prostate Cancer Cell line

Engineering of CNC-based hydrogel nanoparticles with controlled concentrations to enhance the selectivity and efficacy of cisplatin in AGS cells

Studying the molecular and physiological mechanisms of glycine betaine accumulation in order to improve tolerance to abiotic stresses in plants

Bladder cancer treatment with vaccines

REVIEW SPERM DNA BREAKAGE ININDIVIDUALS WITH OLIGOASTHENOSPERMIA AND ITS EFFECT ON ICSI AND EMBRYO FORMATION IN THE INFERTILITY VENTER ROUYESH KARAJ

Comparison of Cryopreservation Methods on Sperm Viability in Domestic Animals: A Systematic Review and Meta-Analysis

Molecular and Cellular Basis of Human Cancer

The effect of pentoxifylline on human sperm parameters and DNA integrity– An in-vitro study

Increasing the Production of Plant Secondary Metabolites Using Biotic Elicitors

Protective Effect of Vitamin E on Sperm Parameters in Adult Rat Treated with Para-nonylphenol

Human sperm cryopreservation update in treatment of infertility: a review study

Evaluation of Curcuma longa L. and bovine ghee effects on experimental stomach ulcers healing in rat

Apoptosis Will be Increased after 2h Incubation of Human Sperm at 37°C

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