t cells

Destruction of dopaminergic neurons causes diseases. Various compounds with neuroprotective and antioxidant properties have been identified, including Hesperidin (HES) and Auraptene (AUR). We aimed in this study to evaluate the in vitro protective effects of these compounds in SH-SY5Y neuroblastoma cell line against the induced neurotoxicity of 6-hydroxydopamine (6-OHDA).

The MTT test to assess cell viability was used. Flow cytometry was conducted for the cell cycle analysis using propidium iodide (PI) stain. The intracellular production of reactive oxygen species (ROS) was assessed using 2, 7′-dichlorofluorescein diacetate (DCFDA) probe and fluorimetry.

Following 6-OHDA treatment, cell viability decreased, and G2/M arrest and ROS levels increased. Our intervention demonstrated that only HES has neuroprotective effects against 6-OHDA-induced toxicity.

HES protects SH-SY5Y cells against 6-OHDA-induced neural damage via inhibiting G2/M arrest, reducing the amount of ROS, and increasing cell viability. However, the different effects and more precise mechanisms are still unknown, and requires new research on animal and human models.

The use of cryoablation for colorectal liver metastases (CLM) remains limited and controversial. This study aimed to investigate the antitumor immune response following cryoablation combined with granulocyte-macrophage colony-stimulating factor (GM-CSF) treatment in a CLM mouse model.

A CLM mouse model was established using BALB/c mice. The tumor-bearing mice were randomly divided into Control group, GM-CSF group, cryoablation group, and cryoablation + GM-CSF group. Tumor size, survival time, dendritic cells (DCs) count, serum cytokine levels (IL-4, IFN-γ), and the Th1/Th2 ratio (IFN-γ/IL-4) were compared among the four groups.

The combination of cryoablation and GM-CSF demonstrated synergistic effects, resulting in the smallest tumor lesion, longest mean survival time, and highest DC count on day 21 post-treatment compared to other groups. Both cryoablation alone and combined with GM-CSF significantly increased serum IFN-γ levels and suppressed IL-4 levels on day 21 compared to pre-treatment levels (P<0.05). Notably, the combination of cryoablation and GM-CSF significantly elevated the Th1/Th2 ratio (P<0.05).

Combining cryoablation with GM-CSF treatment holds promise for CLM management. It exhibits increased DC infiltration within the tumor microenvironment, enhanced immune responses, and prolonged survival in tumor-bearing mice.

Lack of matured oocytes after ovarian stimulation for in vitro fertilization (IVF) is one problem affecting the number of good-quality embryos. In vitro maturation (IVM) rescue is one way that can be done to rescue the immature oocytes retrieved after ovarian stimulation. It has been proven that cumulus cells (CCs) play an essential role in the growth and development of oocytes. However, their role in immature oocytes undergoing IVM rescue still needs to be improved. To assess the role of CCs in immature oocytes after controlled ovarian stimulation (COS) that underwent IVM rescue followed by intra cytoplasmic sperm injection (ICSI).

A retrospective study consists of 200 immature oocytes with normal morphology obtained from IVF patients who had undergone ovarian stimulation. They were randomly divided into a control group (complete denudation, n=100) and a study group (non-denudation, n=100). Both groups then underwent IVM rescue for 24 hours. Oocytes that mature through IVM rescue were followed by ICSI. Our primary outcome is the number of good-quality embryos. Other intermediate outcomes such as oocyte maturation rate, fertilization rate, embryo cleave and rate, and embryo quality were also analyzed.

The two groups had no significant difference in oocyte maturation rate 71 vs. 73% (P=0.875). However, significant differences were found in the fertilization rate 38.1 vs. 61.6% and embryo cleave rates 37.1 vs. 62.2% (P=0.006 and 0.005, respectively). In addition, immature oocytes that underwent IVM rescue with attached CCs produced more good embryos compared to IVM rescue of denuded oocytes 20 vs. 35.7% (P=0.003).

Current research indicates that IVM rescue of immature oocytes with attached CCs may produce more good-quality embryos.

Studies have shown that embryonic stem cells can differentiate into germ cells either spontaneously or directed by the defined factors or by applying a co-culture method or conditioned medium of stromal cells.

Here, Yazd4 (human embryonic stem cells [hESC] line; 46, XX) was used to evaluate the effect of human cumulus cells conditioned medium (CCCM) on in vitro germ cell production and development.

In this experimental study 2 different complete media were applied, Dulbecco’s modified eagle’s medium + 20% fetal bovin serum and embryoid body (EB) medium (KnockOut serum replacement- hESC without basal fibroblast growth factor). Thus, there are 2 control groups for spontaneous differentiation (SD) (SD-EB and SD-DM) and 2 types of CCCM (CCCM-DM and CCCM-EB) as test groups. 40% dilution of the conditioned medium was applied according to previous reports.

Reverse transcription-quantitative polymerase chain reaction and immunofluorescent staining assays revealed that between groups with similar complete medium, CCCM provides a better condition for in vitro germ cell differentiation from hESCs. Moreover, a comparison between the 2 types of complete media showed that EB medium is more supportive than Dulbecco’s modified eagle’s medium +20% fetal bovin serum.

CCCM with EB medium as a complete medium provides a better condition for in vitro germ cell differentiation from hESCs.

This article has been extracted from Ph.D. Thesis. (Somayyeh Sadat Tahajjodi)

Cancer remains the leading cause of death globally, with breast cancer being the foremost cause among women and lung cancer ranking second for both women and men.

This study aimed to identify the metabolomic content of Coleus amboinicus leaves and evaluate their anticancer activities against breast and lung cancer cells, thereby providing insights into potential alternative treatments for these cancers and initiating research on active isolates from C. amboinicus leaves.

The research methodology involved maceration using ethanol, followed by multistage partitioning with solvents nhexane, chloroform, and ethyl acetate. Phytochemical screening was performed using standard reagents to detect the presence of alkaloids, phenolics, polyphenols, flavonoids, steroids/triterpenoids, and saponins. Metabolomic profiling was conducted using LC/HRMS, and the anticancer activities against lung cancer cells (A549) and breast cancer cells (MCF-7) were assessed using the MTT assay.

The results showed that the C. amboinicus extract contains various secondary metabolite groups such as alkaloids, phenolics and polyphenols, flavonoids, steroids, triterpenoids, and saponins.

The diverse metabolomic profile of the C. amboinicus leaf extract demonstrated potential activity against cancer, as evidenced by in vitro tests on lung (A549) and breast (MCF-7) cancer cells. C. amboinicus leaf extract shows promise as an active ingredient in the prevention and alternative natural treatment of lung and breast cancer. Further research and testing, both in vivo and clinically, are warranted.

The incidence of colorectal cancer is increasing globally. Daunorubicin (DNR), an anthracycline antibiotic, is effective against various cancers. The PI3K/AKT/mTOR signaling pathway is crucial in regulating cell growth and cancer growth.

This study aims to evaluate the effects of liposomal daunorubicin (Lip-DNR) on cell proliferation and cell death induction in HCT116 cells compared to free daunorubicin.

Lip-DNR was synthesized, and its shape and size were analyzed using FE-SEM imaging. HCT116 cells were treated with Lip-DNR concentrations of 0 (control), 0.125, 0.25, 0.5, 1, and 2 μm for 48 hours to determine the IC50. The effects of free (0.5 μm) and liposomal DNR (IC50 of 0.43 μm) on PI3K mRNA levels were assessed using real-time PCR. The cell cycle was analyzed by flow cytometry.

FE-SEM imaging showed that the liposomes are spherical and range from 50 - 100 nm in size. Lip-DNR induced cell death in HCT116 cells in a dose-dependent manner, with 0.5 μm Lip-DNR causing more cell death than an equivalent concentration of free DNR. Analysis of PI3K gene expression showed that DNR decreases PI3K gene transcription in HCT116 cells, with Lip-DNR having a more substantial effect than the free form. Both forms reduced the proportion of G2/M phase cells, but Lip-DNR was more effective at inhibiting cell proliferation in HCT116 cells.

DNR inhibits the proliferation of HCT116 cells by downregulating PI3K gene expression and enhancing cell death, with the liposomal form demonstrating stronger effects than the free form.

The potential application of extracellular vesicles (EVs) in regenerative and personalized medicine has attracted substantial interest in recent years, highlighting the need for standardized protocols for their administration in preclinical and clinical settings. EVs, which play critical roles in intercellular communication and have significant therapeutic potential, have prompted extensive research and advancements in their clinical applications. However, the rapid evolution of this field has also revealed variability in how EVs are isolated, characterized, and used across different studies. Over the past decade, organizations such as the International Society for Extracellular Vesicles (ISEV) and the International Society for Cell and Gene Therapy (ISCT) have actively worked to address these challenges by proposing frameworks for standardizing EV-related research. As the clinical evaluation of therapeutic EVs becomes increasingly commonplace, there is a need for practical guidelines and assessment tools that can aid in evaluating their efficacy and safety. In this context, we propose a comprehensive checklist designed to guide researchers and clinicians in considering critical aspects when designing and conducting biomedical and clinical studies involving EVs. This checklist aims to enhance the standardization of trials and therapeutic procedures, ensuring that clinical reports are prepared with adequate detail. By controlling reproducibility and transparency in research, we believe that our proposed guidelines will contribute significantly to advancing the application of EVs in clinical practice.

In recent years, scientists have focused on the use of natural ingredients, including herbal extracts, in the treatment of breast cancer.

This study aimed to evaluate the cytotoxic effects of the aqueous extract of sweet pomegranate seed (AESPS) and vitamin D on MCF-7 breast cancer cells and human fibroblast cells for the first time.

MCF-7 and fibroblast cells were cultured in 96-well plates at a density of 10 4 cells/well and treated with different concentrations of AESPS and vitamin D (25, 50, 100, and 200 mg/mL). Cell viability was assessed using methylthiazol tetrazolium (MTT) assays at 24, 48, and 72 hours. The data were analyzed using statistical tests with a significance level of 0.05.

The results showed that AESPS inhibited the viability of MCF-7 cells, especially at a concentration of 200 mg/mL for 24 hours. In contrast, 200 mg/mL of vitamin D resulted in an increase in cell death of MCF-7 cells after 48 hours. Additionally, AESPS did not cause any significant toxicity to fibroblast cells, while vitamin D caused approximately 30% toxicity to fibroblast cells after 48 hours.

Therefore, this herbal extract has a significant cytotoxic effect on MCF-7 cancer cells, even more so than vitamin D, and a non-cytotoxic effect on human fibroblast cells, significantly less than vitamin D. Aqueous extract of sweet pomegranate seed could be utilized in the treatment of breast cancer with further research in animal models and, eventually, clinical trials.

Context: 

Recent advances in induced pluripotent stem cells (iPSCs), CRISPR-Cas9 gene editing, nanotechnologies, and artificial intelligence have revolutionized regenerative medicine (RM) as a transformative field for tackling difficult medical problems. These breakthroughs promise specific treatments, proper restoration of tissue function, and substantial improvements in the quality of life for patients whose ailments cannot yet be cured.This review explores cutting-edge advancements in RM platforms such as stem cell therapy, gene editing, 3D bioprinting, and nanotechnology. The study also aims to shed light on the challenges of clinical translation and policy implications, which are crucial for fostering sustainable and progressive advances in the discipline. 

Evidence Acquisition: 

This manuscript draws on cutting-edge research on the development and application of RM technologies. It synthesizes data on stem cells, gene therapy, tissue engineering, the in vitro organoid industry, artificial intelligence (AI), and nanotechnology that illustrate therapeutic potential. It also aims to identify ethical, regulatory, and practical hurdles for translating RM from research to clinical practice.

Breakthroughs such as those in iPSC-derived organoids, CRISPR-Cas9 gene editing, 3D bioprinting, and nanostructured materials exhibit significant promise in preclinical and clinical settings. Platforms such as organ-on-chip and AI tools further enhance drug discovery and treatment monitoring, while biomaterials and scaffold-based approaches enhance tissue repair and regeneration. Nevertheless, despite these advances, challenges persist regarding scale-up, safety, and ethical considerations.

Innovations in RM represent a paradigm shift from purely symptomatic treatments to restorative therapies. Successful integration of RM into clinical practice will require multidisciplinary collaborative work, imposition of rigorous safety protocols, and enabling regulatory frameworks. Addressing these challenges would enable RM to realize its true potential as a foundation for 21st-century healthcare.

Earlier studies have highlighted the involvement of miRNA146a in tumor suppression indicating its potential to inhibit the progression of diffuse large B-cell lymphoma (DLBCL).

To identify programmed death-ligand 1 (PD-L1) as a candidate for further research, as it plays a key role in regulating immune checkpoints in cancer and is associated with the involvement of miRNA146a in immune regulation and the response to inflammation.

The expression of miR-146a and PD-L1 in DLBCL cells was detected using qPCR analysis. Subsequently, DLBCL cells (OCI-Ly-3 and OCI-Ly-7) were treated with either the miR-146a mimic or a blank plasmid. To assess immune evasion, DLBCL cells were cocultured with peripheral blood mononuclear cells, CD8+ T cells, or cytokine-induced killer cells. Furthermore, the target gene of miR-146a was predicted and validated.

Compared to the normal B-cell line (NCB), the level of miR-146a was significantly lower in DLBCL cells. Additionally, overexpression of miR-146a significantly reduced DLBCL viability, invasion, and immune evasion while simultaneously promoting apoptosis. Our findings also confirmed that miR-146a targeted PD-L1. Finally, the upregulation of PD-L1 notably reversed the tumor suppressive effects of miR-146a on DLBCL.

Our study indicates that miR-146a inhibits the progression of DLBCL by enhancing antitumor immunity through the targeting of PD-L1. The therapeutic potential of this miRNA in lymphoma is highly desirable.

COVID-19 (2019) clearly demonstrates an imbalanced immune response. Variations in the function and subtypes of dendritic cells (DCs) may have effects on immune responses in COVID-19 patients and contribute to immunopathology.

To assess the phenotype and frequency of Plasmacytoid dendritic cells (pDCs), Conventional DCs (cDCs), and double-positive DCs in COVID-19 patients admitted to the ICU and non-ICU compared to the healthy control group.

The study included 10 healthy individuals and 25 COVID-19 patients. In the second week of their illness, Peripheral blood mononuclear cells (PBMCs) were isolated from the patients and labeled with targeted antibodies for HLA-DR, CD123, and CD11c. The samples were then analyzed using flow cytometry. The COVID-19 patients were divided into two ICU and non-ICU groups and were closely monitored throughout the study.

In comparison to healthy controls, COVID-19 patients exhibited a significantly lower pDCs ratio (P=0.04). Patients were categorized into two groups: (A) the ICU group (n=11; 44%) and (B) the non-ICU group (n=14; 56%). The frequency of pDC was significantly lower in ICU patients than in non-ICU patients (P<0.01). Although not statistically significant, ICU patients had a lower frequency of cDCs and double positive DCs compared to non-ICU patients. Additionally, a significant association between the age of COVID-19 patients and cDC levels was observed (p=0.049).

SARS-CoV-2 can evade attacks from the immune response by reducing the number of DCs and suppressing their function of DCs, ultimately resulting in weakened development of both innate and adaptive immunity.

Hepatocellular carcinoma (HCC) is one of the lethal malignancies with a poor prognosis due to metastatic complications. Matrix metalloproteinases (MMPs) and their inhibitors, tissue inhibitors of metalloproteinases (TIMPs), have an important role in metastasis. MicroRNA-21 (miR-21) is significantly overexpressed in nearly all types of human cancers, including HCC. Targeting miR-21 pharmacologically could be a promising therapeutic approach for HCC. 3,4-dihydroxyphenylethanol (DHPE), a phenolic phytochemical compound found in olive, has potent antioxidant and anticancer properties. This study aimed to investigate the effect of DHPE on the expression of miR-21 with genes associated with metastasis (MMP-2, MMP-9, TIMP-1, and TIMP-2) and their correlation with miR-21 in HepG2 cells.

This experimental study had four groups, including a control, and three groups of treatment with different concentrations of DHPE (50, 100, and 150 µM) for 24 hours. The expression levels of genes were determined by RT-qPCR.

The results showed that the treatment of cells with DHPE significantly reduced the expression of miR-21, MMP-2, MMP-9, and TIMP-1 but increased TIMP-2 compared to the control group; additionally, there was a negative correlation between miR-21 and TIMP-2 but a positive correlation between miR-21 with MMP-2, MMP-9, and TIMP-1.

The results showed that DHPE, likely by reducing the expression of miR-21, can increase TIMP-2 and reduce MMP-2, MMP-9, and TIMP-1 gene expression and may play a role in inhibiting cell migration in HepG2 cells.

Extracellular vesicles (EVs) are cell-derived vesicles that play a critical role in host-pathogen interactions, facilitating intercellular communication and transporting both pathogen- and host-derived molecules during infection spread. To regulate their environment, for instance, by modulating innate and adaptive inflammatory immune responses, pathogens may alter the composition of EVs produced by infected cells. Gastric cancer is one of the leading causes of cancer-related deaths worldwide, and Helicobacter pylori infection is considered a significant risk factor for its development. This cancer is characterized by significant inflammation mediated by EVs generated from infected host cells. H. pylori contributes substantially to inflammation, promoting disease progression. Moreover, H. pylori produces and releases vesicles known as outer membrane vesicles (H. pylori-OMVs), which contribute to the shrinkage and cellular transformation of the gastric epithelium. Although the vacuolating cytotoxin A (VacA) plays a critical role in pathogenesis, its association with EVs in H. pylori has not been previously addressed. Understanding the roles of extracellular vesicles and VacA during H. pylori infection—whether they benefit the host or the pathogen—could pave the way for new treatment approaches. This review briefly discusses the role of VacA and extracellular vesicles in the growth and pathogenesis of H. pylori.

Background and purpose:

Intracellular delivery is crucial in biological and medical studies. Although many molecular tools have been created for cell-based gene therapies, it remains challenging to introduce external molecules into cells. As one of the most popular non-viral transfection methods, electroporation induces transient pores in the cell membrane by applying an external electric field. Unsatisfactory transfection efficiency and low cell viability are the major drawbacks of electroporation. To overcome these issues, the current study investigated the effect of urea on electroporation-mediated transfection efficiency.

Experimental approach:

Three voltages of electroporation, including 100, 120, and 140 V, and 3 concentrations of urea buffer, including 0.25%, 0.5%, and 1% W/V, were considered as variables in this study. The HEK-293 cell line was used for transfection, and green fluorescent protein (GFP) expression was evaluated using flow cytometry and fluorescence microscopy.

Findings/Results:

The results showed that the combination of electroporation and urea increased electroporation efficacy, but the effect depended on voltage and urea concentration. When different concentrations of urea were added to HEK-293 cells at a voltage of 100 V, the number of cells transfected by pEGFP-N1 increased (from 12.3 ± 0.2% in untreated cells to 17.35 ± 0.55%, 23.3 ± 0.3%, and 14 ± 0.1% at urea concentrations of 0.25%, 0.5%, and 1% W/V, respectively). The electroporation buffer containing 0.5% W/V urea showed the highest EGFP expression (23.3 ± 0.3%) and high cell viability (over 90%).

Conclusion and implications:

This research offers a new perspective for improving gene transfection efficiency once electroporation is utilized.

Cell-based therapy has shown promising outcomes in the treatment of cerebral palsy (CP). However, there is noconsensus on a standard therapeutic protocol regarding the source of cells, optimal cell dose, timing and frequencyof cell injections, route of administration, or the use of combination therapy. This lack of consensus necessitates acomprehensive investigation to clarify these crucial yet undefined factors in cell-based therapy for CP patients. In thiscommentary, we discuss and compare the trends in Gross Motor Function Measure-66 following intrathecal injectionof umbilical cord blood mononuclear cells (UCB-MNCs) and umbilical cord tissue mesenchymal stromal cells (UCTMSCs)in children with CP. Our study revealed that MNC injections led to earlier improvements in gross motor function,whereas MSC applications resulted in more sustainable changes. These findings provide key insights into the efficacyof different cell types, which will be beneficial for future studies and for refining cell-based therapy protocols for CPtreatment.

Chronic lymphocytic leukemia (CLL) is the most prevalent hematological cancer, with various medical interventions. In the recent decade, cold physical plasma has become an interesting agent for future cancer therapy. The goal of this study was to see whether cold physical plasma or cold physical plasma-treated liquid (PTL) affected integrin beta 3 (ITGB3) expression, which is hypothesized to mediate an interaction between cancer stem cells and the bone marrow microenvironment, in CLL patients' blood cells. The metabolic activity, cell death pattern, lipid oxidation and ITGB3 gene expression of these treatments was evaluated. Both direct cold physical plasma and PTL exposure enhanced lipid peroxidation in cells of CLL patients, but to a lesser extent in healthy participants. Furthermore, following 48h of cold physical plasma or PTL exposure, the metabolic activity of leukocytes was preferentially reduced in CLL patient leukocytes. In addition, cold physical plasma and PTL treatment elevated ITGB3 mRNA expression in CLL patients' leukocytes compared to untreated and healthy controls. Collectively, our study suggests selective effects of direct cold physical plasma and PTL exposure on blood leukocytes from leukemia patients, but further and more detailed studies are needed to provide additional rationales for such treatment options as future therapy.