Gene, Cell and Tissue
Volume:12 Issue: 2, Apr 2025

Lentivirus infection significantly impacts gene expression in host cells, including the regulation of housekeeping genes, which are essential for normalization in quantitative reverse transcription PCR (RT-qPCR). This normalization reduces measurement errors arising from sample quality variations, RNA extraction methods, and experimental conditions. In the context of glioblastoma (GBM) and neuroblastoma (NB) — two malignancies characterized by poor prognoses and limited progression-free survival — it is crucial to identify stable reference genes to ensure accurate gene expression analysis.

Reliable normalization provides more trustworthy insights into the molecular mechanisms underlying tumor progression and may inform the development of targeted therapeutic strategies.

The expression stability of eight housekeeping genes (RPL32, RPS23, GAPDH, 18S rRNA, TUB, ACTB, HPRT, and TBP) in U87 glioblastoma cells and seven genes (RPL32, HPRT, GAPDH, 18S rRNA, TUB, ACTB, and RPII) in SH-SY5Y NB cells was evaluated using RT-qPCR. Gene stability was analyzed using four statistical tools: GeNorm (pairwise variation-based ranking), NormFinder (model-based variance estimation), BestKeeper (standard deviation and CV analysis), and RefFinder (integrative ranking). Each experiment was performed in three biological replicates and analyzed in technical triplicate to ensure statistical robustness.

In SH-SY5Y cells, ACTB, RPL32, and RPII were consistently identified as the most stable reference genes across GeNorm, NormFinder, and BestKeeper analyses, while TUB was ranked as the least stable. In U87 cells, GeNorm ranked RPS23/HPRT as the most stable, NormFinder favored TUB/GAPDH, and BestKeeper prioritized ACTB/RPL32. Although slight differences in the gene rankings were observed among the different statistical tools, the overall selection of the most stable reference genes remained consistent. RefFinder’s integrative analysis resolved these discrepancies, identifying 18S (M = 0.18) and GAPDH as the most stable genes, and ACTB/HPRT as the least stable genes in U87. In SH-SY5Y, ACTB and RPL32 (M = 0.22) were the most stable genes, and TUB was the least stable (M = 2.45).

This study provides a framework for reliable gene expression analysis in lentivirus-infected models. Our findings highlight the context-dependent stability of housekeeping genes, necessitating validation in diverse experimental settings (e.g., alternative viruses, primary cells) to ensure broader applicability. By emphasizing rigorous normalization, this work enhances reproducibility in gene expression studies and advances translational research in viral oncology and neuro-oncology, particularly for glioblastoma and NB therapeutics.

The damage to agricultural products caused by fungal diseases accounts for approximately 12% of global production, with a higher impact in developing countries. One particularly destructive disease that inflicts significant damage on vegetable crops and greenhouses annually is seedling death and bush dieback disease, caused by the Phytophthora fungus. Currently, producers often rely heavily on chemical fertilizers and pesticides to enhance productivity and product quality. However, the use of some synthetic pesticides has recently been banned due to their high toxicity and long preharvest interval (PHI) periods. Biological control of plant diseases and pests is the optimal solution to mitigate the damages caused by synthetic pesticides.

This study aimed to evaluate the in vitro antifungal efficacy of different extracts derived from the aerial shoot of Indigofera tinctoria against plant pathogenic fungi.

The aerial shoots of the Indigo plant were collected during the flowering stage from the medicinal plants collection of the Institute of Agricultural Research, Agriculture Institute Research, Institute of Zabol, Zabol, Iran. Ethanolic, methanolic, n-hexane, and acetone extracts were prepared by cold maceration with a ratio of 1:10. The antifungal effects of these extracts were investigated by measuring the diameter of halo growth inhibition under in vitro conditions. The LSD test was used to compare data means.

Our results showed that the best growth-inhibiting effect was observed with the ethanolic extract at a concentration of 300 μg/mL (2 mm) after 24 hours of incubation, whereas the diameter of colonies in the control was 27 mm, confirming a 92.5% inhibition of fungal growth. After 48 hours of incubation, the growth of Phytophthora drechsleri mycelium was clearly evident. In fact, the growth rate nearly doubled within 48 hours, but then at 73 hours, the mycelium grew at a slower rate. The inhibitory effect of the indigo extract was dose-dependent; as the extract concentration increased, the inhibitory effect became stronger.

According to these results, the in vitro treatments were more significant when the plant extract concentration was higher. These findings could be promising for the production of natural fungicides.

Tendons are subjected to mechanical stress and strain during daily activities and physical exercise, which makes them susceptible to injury.

This randomized trial evaluated the effects of a 4-week combination of training and celastrol on the healing of Achilles tendon ruptures in rats. It assessed cytokine levels, gene expression, and histological changes.

Rats with Achilles tendon ruptures were randomly assigned to several treatment groups: Sham, negative control, training, celastrol, and a combination of training and celastrol. Eligibility criteria included age, weight, and the absence of pre-existing tendon injuries. The assessments included enzyme-linked immunosorbent assay (ELISA) to measure cytokine levels [bFGF, IGF1, cyclooxygenase 2 (COX2)], real-time polymerase chain reaction (RT-PCR) to analyze gene expression (scleraxis, periostin, MMP9, collagen I), and histological analysis using Masson's trichrome staining to evaluate collagen organization.

The training-celastrol group exhibited the highest levels of bFGF, IGF1, and COX2, indicating a synergistic effect on tendon healing compared to the sham and negative control groups, with effect sizes of 0.85 (95% CI: 0.70 - 1.00). Additionally, RT-PCR results showed that the training-celastrol group had the highest expression of scleraxis, periostin, MMP9, and collagen I, suggesting enhanced tendon repair and remodeling. Histological analysis revealed that the training-celastrol group displayed the most organized collagen fibers, whereas the sham group showed regular organization, and the negative control group exhibited disorganized fibers.

The study concluded that combining aerobic training and celastrol significantly enhances tendon healing by improving cytokine levels, gene expression, and collagen organization. Outcome measures were assessed at baseline, mid-treatment (week 2), and post-treatment (week 4) to evaluate temporal changes. These findings highlight the effectiveness of these therapeutic strategies in promoting tendon repair and maintaining structural integrity.

 Colorectal cancer (CRC) is a leading cause of cancer-related deaths globally. Its progression is influenced by various molecular factors, including the dysregulation of microRNAs (miRNAs). MiR-373 is an oncogenic miRNA implicated in the development of CRC, but its precise role in tumor progression and metastasis remains under investigation.

This review aims to evaluate the role of miR-373 in CRC progression, focusing on its impact on key cellular processes such as proliferation, migration, and invasion. Additionally, the review explores the potential of miR-373 as a prognostic biomarker and therapeutic target in CRC.

A systematic search was conducted using databases such as PubMed, Scopus, and Google Scholar to identify studies published from 2000 to 2025. The review includes studies that investigate miR-373 expression, its regulation of tumor suppressor genes, and its involvement in oncogenic signaling pathways, particularly those linked to CRC progression.

MiR-373 is often overexpressed in CRC tissues, promoting tumor growth by regulating critical cellular processes. It suppresses tumor suppressor genes like PTEN and TP53INP1, resulting in uncontrolled cell proliferation, reduced apoptosis, and enhanced invasion. Higher miR-373 levels are associated with advanced disease stages, metastasis, and poor clinical outcomes, suggesting its potential as a prognostic marker and therapeutic target.

MiR-373 contributes significantly to CRC development and progression. Its upregulation is linked to increased tumor aggressiveness, metastasis, and resistance to therapy, making it a promising candidate for early detection and targeted therapies in CRC. Further studies should focus on modulating miR-373 expression to improve clinical outcomes.

Natural dyes derived from plants have gained significant attention due to their safety compared to chemical dyes. These dyes possess antibacterial properties and have minimal environmental impact.

A systematic literature search was conducted using PubMed, Scopus, Embase, Cochrane, Web of Science, and Google Scholar databases. The search utilized all available MeSH terms related to medicinal plants, dyes, wool, and antimicrobial properties of plants, covering the period from 2009 to 2020.

The findings indicate that numerous plants possess both coloring and antimicrobial properties, although they are currently utilized in limited quantities.

The present study aims to introduce plants with antimicrobial and coloring properties for further study.

One of the most common techniques in assisted reproductive technologies (ART) is in vitro embryo production (IVEP), the success of which depends on the intrinsic quality of oocytes and the composition of the culture medium.

This study investigates the effects of gonadotropin hormones (such as eCG and hCG) along with the antioxidant quercetin on the maturation of ovine oocytes, as well as the impact of different media on the expression of Bcl2 and Bax genes in blastocysts.

After washing the oocytes, cumulus-oocyte complexes (COCs) that exhibited three or more layers of cells and homogeneous cytoplasm were selected and matured in BO-IVM, TCM (TCM-199 + 10% FBS + 10% ovine follicular fluid + 5 mg/mL FSH + 1 mg/mL estradiol-17β + 0.81 mM sodium pyruvate + 50 mg/mL gentamicin sulfate), and TCM+ (TCM + 20 µg/mL eCG + 5 µg/mL hCG + 15 µg/mL quercetin) media for 24 hours at 38.5°C, 20% O2, and 5% CO2. After maturation, oocytes were fertilized with frozen ram semen, and presumed zygotes were cultured in BO-IVC under uniform conditions. Embryo development occurred at 38.5°C in a humid atmosphere with 5% CO2, 5% O2, and 90% N2.

The results indicated that the BO-IVM medium had a significantly higher mean percentage of zygotes, morulae, blastocysts, and hatched blastocysts compared to TCM+ and TCM (P < 0.05). Moreover, blastocyst formation and hatched blastocysts were significantly more frequent in TCM+ than in TCM (P < 0.05). However, the BO-IVM medium demonstrated a significant improvement compared to other maturation media. Gene expression analysis showed no significant difference in Bcl2 expression among the groups, while Bax expression was significantly higher in TCM (P < 0.05). Overall, the BO-IVM medium was identified as the best option for optimizing oocyte maturation and producing high-quality blastocysts in ovine, with TCM+ yielding better results than the TCM medium without hormonal and antioxidant compounds.

The results of this study showed that the addition of the antioxidant quercetin and the growth factors eCG and hCG improved the performance of the TCM medium. However, further research is needed to enhance the performance of the TCM maturation medium compared to the commercial BO-IVM medium.

Insulin-like growth factor 1 (IGF-1) is an important regulator of growth, amino acid (AA) elongation, glucose metabolism, DNA synthesis, protein synthesis, and the proliferation and differentiation of various cell types. Therefore, IGF-1 may affect intestinal morphology by increasing nutrient uptake into intestinal enterocytes and promoting cell proliferation.

The present study aimed to evaluate the effects of different levels of branched-chain amino acids (BCAAs) (L-valine, L-leucine, and L-isoleucine) in low crude protein (CP) diets on intestinal morphology and IGF-1 gene expression in broiler chickens.

A total of 480 one-day-old male and female broilers of the Ross 308 strain, with an average weight of 42.75 ± 0.47 g, were used. This experiment was carried out as a 3 × 2 factorial in the form of a completely randomized design with 6 treatments and 4 replications. The experimental diets included three levels of BCAA (0%, 10%, and 20% higher than the standard) and two levels of CP (standard or 10% lower than the standard).

Using 20% BCAA in the diet increased the villus height to crypt depth (VH/CD) ratio in the duodenum, jejunum, and ileum compared to the standard level of BCAA (P < 0.05). Reducing CP by 10% significantly lowered the VH/CD ratio in the duodenum, jejunum, and ileum (P < 0.05). The expression of IGF-1 mRNA in liver tissue was higher in the groups containing 10% BCAA than in the standard BCAA groups, regardless of the CP level.

In general, BCAA supplementation could be beneficial for improving intestinal morphology and IGF-1 gene expression in broiler chickens on low CP diets.