Biomedical Research and Therapy

Dysregulation of miR-124-3p and STAT3 in Hypothyroidism: Implications for Diagnostic and Therapeutic Biomarkers

Akshay Kumar Anand Kumar — 2025-05-31

Background: Hypothyroidism is a clinical condition caused by decreased thyroid hormone (T3 and T4) levels, affecting ~5% of the population. However, its molecular mechanisms remain poorly understood. Recent studies implicate microRNAs (miRNAs) in disease-related cellular processes. This study aimed to identify novel miRNAs associated with hypothyroidism using publicly available genomic databases, with the goal of validating one candidate miRNA and its target gene in patients to evaluate their potential as diagnostic and therapeutic biomarkers.

Methods: We used bioinformatic tools (NCBI, TargetScan, miRBase) to identify candidate miRNAs, selecting miR-124-3p for further analysis. The secondary structure of miR-124-3p was predicted using RNAfold. We quantified expression levels via qPCR (quantification cycle, Cq; melt curve analysis). Similarly, we analyzed expression of Signal Transducer and Activator of Transcription 3 (STAT3), a predicted target of miR-124-3p.

Results: Secondary structure prediction revealed that miR-124-3p has a minimum free energy of -35.50 kcal/mol. miR-124-3p was downregulated and STAT3 was upregulated in hypothyroid patients, suggesting their involvement in pathogenesis.

Conclusion: Our computational and experimental data indicate that miR-124-3p is dysregulated in hypothyroidism and may serve as a diagnostic, prognostic, and therapeutic biomarker. Altered expression of miR-124-3p and its target STAT3 provides insights into hypothyroidism pathogenesis. Further studies are needed to elucidate their roles. STAT3 also shows potential as a biomarker target.

Nipah Virus: Transmission Dynamics of a Zoonotic Outbreak and Therapeutic Challenges

Venkatakrishnan Kiran — 2025-05-31

Nipah virus (NiV) is a deadly zoonotic virus that has caused multiple outbreaks since it was first identified in Malaysia in 1998. It is primarily transmitted by fruit bats and can spread among humans, leading to severe neurological and respiratory complications. With fatality rates ranging from 40% to 75%, NiV poses a serious public health threat, and there are currently no approved treatments or vaccines. The virus enters human cells via ephrin-B2 and ephrin-B3 receptors, causing extensive harm while evading the immune system, which complicates treatment efforts. Environmental changes, including deforestation and increased human–wildlife contact, have heightened the likelihood of NiV outbreaks. Preventing future outbreaks necessitates early detection, strict biosecurity measures, and concerted global efforts to develop antiviral therapies and vaccines. This review comprehensively examines NiV's virology, transmission dynamics, clinical manifestations, and the latest advances in therapeutic care, encompassing One Health strategies and preventive measures.

Unraveling Common Stem Cell Sources and Key Reporting Parameters in Studies Related to Stem Cell-Derived Red Blood Cells: A Review

Siti Salmah Noordin — 2025-05-31

Introduction: Blood transfusions are essential for maintaining oxygen delivery to tissues in cases of severe blood loss. However, challenges such as limited donor availability, short storage lifespans, blood-type incompatibility, and infection risks necessitate alternative solutions. Stem cell-derived red blood cell (RBC) substitutes offer a promising approach to address these limitations. Multiple stem cell sources, including embryonic stem cells (ESCs), hematopoietic stem cells (HSCs), and induced pluripotent stem cells (iPSCs), have been explored for RBC generation. ESCs pose ethical and immunogenicity concerns; HSCs exhibit limited proliferation potential and variable outcomes; iPSCs face safety, standardization, and scalability challenges for RBC generation. Despite significant research in this area, no comprehensive mapping of the evidence exists. This scoping review aims to systematically map the literature on stem cell-derived RBC substitutes and identify key reported parameters.

Methods: This study follows the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Statement for Scoping Reviews (PRISMA-ScR) framework for scoping reviews, encompassing five key stages: defining research questions, identifying relevant studies, selecting eligible articles, data charting, and summarizing findings. A systematic search was conducted using the PubMed and Scopus databases. From 11,074 identified articles, 15 studies met the eligibility criteria. Extracted data focused on stem cell sources, culture conditions, RBC maturation (enucleation rate and hemoglobin composition), and expansion efficiency.

Results: The analysis revealed that ESCs were the most frequently utilized stem cell source, followed by HSCs and iPSCs. HSCs demonstrated the most favorable outcomes, with faster culture times (fewer than 21 days) and higher enucleation rates, ranging from 50% to 98% in some studies. ESCs exhibited higher RBC yields but showed lower enucleation efficiency. In contrast, iPSCs had the lowest enucleation rates, indicating challenges in their use for RBC generation. Key culture parameters, including cytokine supplementation, oxygen tension, and differentiation protocols, significantly influenced RBC yield and maturation.

Conclusion: Stem cell-derived RBCs represent a viable alternative to conventional blood transfusions, offering an unlimited source of RBCs while addressing donor-related challenges. Among the examined stem cell types, HSCs demonstrated the most promising characteristics in terms of culture efficiency and enucleation rates. This review provides a comprehensive overview of essential parameters for advancing RBC generation and serves as a valuable resource for future research in the development of stem cell-based blood substitutes.

Non-Invasive Prenatal Testing: Advances, Applications, and Limitations in Prenatal Screening

Ting Yu — 2025-05-31

Non-invasive prenatal testing (NIPT) is a prenatal screening technology based on the analysis of cell-free fetal DNA (cfDNA) detected in maternal peripheral blood. It offers high detection efficiency for common chromosomal aneuploidies, such as trisomy 21 (T21), trisomy 18 (T18), trisomy 13 (T13), and sex chromosome aneuploidies (SCA). Additionally, NIPT has expanded to include the screening of subchromosomal microdeletions and microduplications, single-gene genetic diseases, and has even demonstrated certain diagnostic value for placental-derived complications during pregnancy. However, some of the problems it presents, such as technical limitations and ethical or psychological issues, cannot be overlooked. This article reviews the advancements and limitations of NIPT in prenatal screening.

TLR4 Agonist CRX-527 Modulates Intracellular and Inflammatory Cytokine Expression in Lymphoid Tissues of BCG-MSP1C-Immunized Mice

Norlaily Hanifi — 2025-05-31

Introduction: Due to the complexity of the malaria parasite’s life cycle, together with the limited knowledge of its intricate immunological response, this study was conducted. This study aims to evaluate the ability of a Toll-like receptor 4 (TLR4) agonist, CRX-527, to enhance the production of intracellular and inflammatory cytokines in lymphocyte-derived tissues of malaria vaccine candidates.

Methods: A total of forty-eight male Balb/c mice was randomly divided into eight groups (n = 6 per group). Two groups of mice were immunized intraperitoneally with 2 × 10⁶ colony-forming units (CFU) of parent Bacillus Calmette–Guérin (BCG) in 200 µl of 1 × phosphate-buffered saline–Tween 80 (PBS-T80) in the presence or absence of 0.5 mg/kg of CRX-527. Another two groups of mice were immunized with 2 × 10⁶ CFU of Mycobacterium bovis Bacillus Calmette–Guérin expressing the C-terminus merozoite surface protein-1 of Plasmodium falciparum (BCG-MSP1C) in 200 µl of 1 × PBS-T80 in the presence or absence of 0.5 mg/kg of CRX-527. Meanwhile, another two groups of mice were immunized with 200 µl of 1 × PBS-T80, whereas the remaining two groups were immunized with 200 µl of lipopolysaccharide (LPS) in the presence or absence of 0.5 mg/kg of CRX-527, serving as control groups. The lymphoid tissues obtained from the spleen, lymph nodes, and liver were harvested from the euthanized mice and were cultured for 24 hours. Then, the supernatant was collected to detect the production of selected intracellular and inflammatory cytokines using a sandwich enzyme-linked immunosorbent assay (ELISA).

Results: There was a significant increase (p < 0.05) in tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), interferon-gamma (IFN-γ), and interleukin-4 (IL-4) in the presence of CRX-527, with the highest levels observed in the spleen, followed by the lymph nodes and liver from the BCG-MSP1C groups, followed by the parent BCG-immunized groups.

Conclusion: These findings demonstrate that the increased production of several intracellular and inflammatory cytokines by the developed malaria vaccine candidate (BCG-MSP1C) in the presence of CRX-527 plays an important role in triggering innate immune responses in lymphocyte-derived tissues.

Double Vitrification-Warming Cycles Reduce Live Birth Rates in Single Euploid Blastocyst Transfers: A Retrospective Cohort Study

Thuy Tran Thi — 2025-05-31

Objectives: To evaluate the impact of blastocysts biopsied once and vitrified twice on clinical outcomes.

Methods: This retrospective study analyzed 277 single euploid blastocyst transfer cycles conducted at the Center for Assisted Reproduction, 16A Ha Dong General Hospital, from March 2018 to January 2024. Cycles were stratified into two groups: Group BV (biopsied once, vitrified once; n = 207) and Group VBV (biopsied once, vitrified twice; n = 70). Pregnancy outcomes were compared between groups, and a binary logistic regression model identified variables associated with live birth rates.

Results: Group BV demonstrated superior outcomes compared to Group VBV across all metrics: implantation rate (55.6% vs. 37.1%, p < 0.001), clinical pregnancy rate (55.1% vs. 37.1%, p < 0.001), ongoing pregnancy rate (54.1% vs. 35.7%, p < 0.001), and live birth rate (53.6% vs. 35.7%, p = 0.01). The number of vitrification-warming cycles was the only factor significantly associated with reduced live birth rates (OR 1.95, 95% CI 1.01–3.78, p < 0.05).

Conclusions: An additional vitrification-warming cycle significantly reduces pregnancy success in single euploid blastocyst transfers. Patients undergoing PGT-A should be explicitly counseled about the potential decline in success rates if previously vitrified blastocysts undergo repeated warming cycles for diagnostic confirmation.

Impact of Antioxidant Supplementation on In Vitro Growth of Porcine Oocytes Derived from Early Antral Follicles

Lam Chi Thien — 2025-05-31

Introduction: Oxidative stress (OS), a key factor impairing oocyte developmental competence, poses a major challenge in in vitro growth (IVG) systems. While antioxidant supplementation can mitigate OS-driven deficits, combining agents with complementary mechanisms—such as melatonin (Mela) and astaxanthin (Asta)—may offer synergistic benefits. This study evaluates the effects of Mela and Asta, individually and in combination, on oocyte quality, gene expression, and epigenetic markers during IVG.

Methods: Growing oocytes from early antral follicles (EAFs; 1–1.5 mm) were cultured for 4 days in a 3D agarose system. Initial experiments tested Mela (1, 10, 100 µM/mL) to assess survival rates and expression of BMP15, CAT, SOD1, and GPX4 via qPCR. Subsequently, the optimal Mela concentration was combined with Asta (0.05, 0.1, 0.25 µM/mL) to determine synergistic effects. Histone H3 methylation (Me-H3-K4) and global transcriptional activity (nascent mRNA accumulation) were analyzed to evaluate chromatin remodeling and transcriptional dynamics.

Results: Mela supplementation significantly improved oocyte survival compared to controls, with 1 µM/mL Mela yielding optimal results. Combining 1 µM/mL Mela with 0.05 µM/mL Asta further enhanced survival rates and upregulated antioxidant genes (CAT, SOD1, GPX4) and BMP15. This combination also elevated Me-H3-K4 levels and nascent mRNA accumulation, indicating enhanced transcriptional activity and chromatin remodeling. These changes correlated with reduced ROS impact and improved oocyte quality post-IVG.

Conclusion: Co-supplementation with Mela (1 µM/mL) and Asta (0.05 µM/mL) synergistically mitigates OS during IVG by boosting antioxidant gene expression, enhancing epigenetic modifications (Me-H3-K4), and promoting global transcription. This strategy significantly improves oocyte developmental competence, offering a promising approach to optimize IVG systems.