Biotechnic & Histochemistry最新文献

Safeguarding the integrity and functionality of the gastrointestinal system is paramount, given its vulnerability to several detrimental effects. One of the factors that can cause functional disorders is oxidative stress, which can disrupt the homeostasis of intestinal tissue and cause various diseases. In this study, we aimed to evaluate the expression patterns of nuclear factor erythroid 2-like 1 (Nrf1) and nuclear factor erythroid 2-like 2 (Nrf2) transcription factors, which are part of an important cleaning system that protects cells against oxidative stress, in the adult rat small intestine by immunohistochemical means. Six Wistar albino adult rats were used in this study. After taking 5 μm thick sections of the small intestine, immunohistochemistry labeling was performed to investigate the expression of Nrf1 and Nrf2. Both transcription factors were found to exhibit immunopositivity in the duodenum, jejunum, and ileum segments of the small intestine. In the crypt epithelium, Nrf1 and Nrf2 showed similar intensity and distribution of staining, whereas, in the villus epithelium, Nrf2 immunoreactivity was most prominent in the ileum and weakest in the jejunum. Nrf1 exhibited lower staining intensity in the ileum. In the smooth muscle layers and the myenteric plexus, Nrf2 immunopositivity was highest in the duodenum, while it was weaker in the jejunum and ileum. These findings indicate that Nrf1 and Nrf2 display region-specific expression patterns in the small intestine and suggest that these transcription factors may play distinct roles in the regional oxidative stress response of intestinal tissues.

Our study aimed to investigate the antidiabetic, antioxidative, and antiapoptotic effects of liraglutide on the testes of rats with diabetes and ischemia/reperfusion injury. Subjects were divided into three groups: control, diabetes, and torsion groups. Rats with diabetes were further divided into two subgroups such as diabetes and diabetes+Liraglutide groups. The torsion group was divided into three subgroups such as torsion, torsion/detorsion, and torsion/detorsion+Liraglutide groups. Malondialdehyde (MDA), superoxide dismutase (SOD), and testosterone levels were measured from blood samples. Also, testicular tissue samples were examined by light and electron microscopy. Apoptosis was assessed using immunohistochemistry for caspase-3. Degeneration of seminiferous tubules and interstitium was observed in the diabetes, torsion, and torsion/detorsion groups, while Liraglutide treated groups showed normal seminiferous tubules morphology. Elevated levels of apoptosis, i.e. caspase-3, were observed in diabetes, torsion, and torsion/detorsion groups (P < 0.05), whereas Liraglutide treated groups had similar levels of apoptosis as the control group. MDA levels of diabetes, torsion, and torsion/detorsion groups were increased (P < 0.05), while SOD and testosterone levels were decreased (P < 0.05). However, Liraglutide treated groups, SOD, MDA, and testosterone levels were found similar to the control group. In conclusion, Liraglutide positively affects structural changes and hormone levels in diabetes and torsion/detorsion groups.

The present study investigated the protective effect of Aloe vera extract against the toxic effects of doxorubicin (DOX) on mice ovaries cultured in vitro. Female Swiss mice with regular estrous cycle were used. The animals (n = 48) had their ovaries collected and cultured individually in a 24-well plate at 37.5°C in 5% CO₂ for 6 days. The ovaries were culture DMEM⁺ alone or supplemented with DOX (0.3 μg/ml), as well as both DOX and Aloe vera (5%, 10%, 25%, or 50%). After culture, ovaries were fixed for histological analysis (follicle morphology, growth and activation, extracellular matrix (ECM) configuration and stromal cell density), immunohistochemistry (TNF-α expression) or stored at - 80°C to evaluate the levels of mRNA for superoxide dismutase (SOD), catalase (CAT), nuclear factor-erythroid 2-related factor 2 (NRF2) and tumoral necrosis factor-α (TNF-α) by real-time PCR. The results showed that ovaries cultured with DOX had a lower percentage of normal follicles and reduced stromal cell density, but when Aloe vera extract was added to the culture medium, there was a protective effect on the ovarian structure against the deleterious effects of DOX. In addition, ovaries cultured with both DOX and Aloe vera (10 and 25%) had reduced TNF-α immunostaining and increased expression of mRNA for antioxidant enzymes (SOD, CAT, and NRF2). In conclusion, Aloe vera is associated with a protective effect on ovarian follicles and stromal cells against DOX-induced toxicity. Therefore, Aloe vera has great potential to preserve fertility in patients undergoing chemotherapy treatment, which often cause irreversible damage to oocytes.

Breakthrough progress has been made in the molecular mechanism research and clinical application of PD-1/PD-L1 in the regulation of immunosuppression and tolerance mainly in human and mouse fields, but is relatively slow in other species. The eukaryotic expression vectors pECFP-Fc-1 and pEYFP-Fc-L1 for high expression of canine PD-1 and PD-L1 proteins were constructed and transfected into human embryonic kidney 293 T cells. Fluorescence microscopy, laser scanning confocal microscopy, and immunofluorescence technology were used to identify the expression and membrane localization of the target proteins in human embryonic kidney 293 T cells. The binding activity of the target proteins expressed on the model cell was identified by eukaryotic expression vector co-transfection and immunocoprecipitation. The results showed that canine PD-1 and PD-L1 proteins were expressed on the membrane surfaces of their respective positively transfected cells. The cell membrane complex was further analyzed by co-immunoprecipitation technology, PD-L1 protein components were successfully detected in the pull-down complex of canine PD-1 antibody, and the two target proteins expressed in the model cells showed good mutual binding activity. Further research is needed to evaluate high throughput and a reliable method for screening drugs that block the PD-1 and PD-L1 pathway.

One of the most widely used techniques for cancer treatment is radiotherapy. Autophagic pathways allow some cancer cells that are resistant to radiation therapy to survive. Inhibiting autophagy has been shown to improve radiotherapy efficacy in several cancer types. Chloroquine (CQ) is a reasonable choice that has been used for many years to treat malaria and is preferred because of its minimal side-effects. Nevertheless, the effects of coadministration of CQ with radiation on various tissues remain unclear. In this study, it was aimed to understand how CQ, used to increase the effectiveness of radiotherapy, has effects on small intestine tissue alone and together with radiotherapy and what role apocynin (APO) can play with its antioxidant character in these stress conditions. Animals were divided into eight groups. The control group received physiological saline, while the other groups received 8 Gy total body irradiation, 50 mg/kg CQ, and 20 mg/kg APO, alone and in combination. In addition to causing significant histological damage, radiation triggered autophagy and showed antiproliferative and apoptotic effects. CQ administered with radiotherapy (RAD) had antiproliferative effects and did not cause a significant change in apoptosis. Reduced glutathione level, glutathione reductase, glutathione peroxidase, glutathione S-transferase, catalase, superoxide dismutase activities, and total antioxidant status were decreased, while lipid peroxidation, total oxidant status, reactive oxygen species, tumor necrosis factor-α, nitric oxide levels, adenosine deaminase, alkaline phosphatase, trypsin, lactate dehydrogenase, sodium potassium ATPase, xanthine oxidase activities, and protein carbonyl contents were increased in the RAD, CQ, and RAD+CQ groups. Apocynin therapy reversed these effects.

Liver ischemia and ischemia/reperfusion (I/R) injury are common in hepatic resection, trauma surgery, and transplantation and contribute to postoperative morbidity and mortality. This study aimed to investigate the relationship between early histopathological changes due to liver I/R injury and serum levels of perilipin-2 (Plin2) and other oxidative stress biomarkers. Fifty female Wistar albino rats were divided into five groups: Group 1 (control), Group 2 (ischemia for 30 minutes), and Groups 3-5 (ischemia followed by 1, 2, or 3 hours of reperfusion). Intracardiac and arterial blood samples were collected for biochemical analysis, while liver tissue samples were used for histopathological examination. Serum levels of Plin2, ischemia-modified albumin (IMA), total antioxidant status (TAS), and total oxidant status (TOS) were measured. Plin2 levels were significantly lower in the ischemia group compared to others (p < 0.01). The control group had significantly lower IMA, TOS, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) levels (p < 0.01) and lower TAS levels (p < 0.05). Rats with Grade 1 liver injury showed significantly lower Plin2 (p < 0.01) and higher IMA levels (p < 0.05). Reduced serum Plin2 following ischemia suggests its potential as a biomarker for acute liver injury. Further studies are needed to clarify its role across different reperfusion durations.

Hematoxylin and eosin (H&E) stain, the routine histological staining method, might not accurately represent the composition and properties of hard tissues. This limitation necessitates the use of advanced diagnostic methods. Methylene blue-acid fuchsin (MB-AF) and modified Gallego's iron fuchsin (MG) stains are useful for differentially staining hard tissues, such as teeth, bone, and pathological calcifications. This study aimed to evaluate and compare the efficacy of MG and MB-AF stains in lesions composed of calcified tissues. A total of 30 histopathologically confirmed cases of various lesions composed of calcified tissue, including compound odontoma (6), ossifying fibroma (6), osteosarcoma (6), osteomyelitis (6), and fibrous dysplasia (6), were chosen. Each lesion had three sections stained with MB-AF, MG, and H&E. Wilcoxon signed rank test was employed for statistical analysis. Dentin, cementum, and bone showed light green, red, and deep green hues, respectively, when stained with modified Gallego's stain. A statistically superior intensity and contrast (p < 0.05) was observed in the MG-stained sections as compared to those stained with MB-AF. MG stain demonstrates superior potential as a special stain for calcifications compared to MB-AF, particularly in terms of the contrast between hard tissues and the surrounding stroma in lesions composed of calcified tissue.

Although Prunus persica var. nucipersica (PPN) exerts antinociceptive, antipyretic, antitumor, anti-allergic, and anti-inflammatory activities with various benefits for human health, the efficacy of the leaf extract from PPN (PPNLE) for wound healing has not been studied yet. In this study, we found that PPNLE clearly affected the scavenging of in vitro free radicals for wound healing. Furthermore, in fibroblast cells, PPNLE significantly resulted in the increased mRNA and protein expressions of wound healing factors, and induced migration of fibroblast cells. In addition, vascular endothelial growth factor (VEGF) secreted from fibroblasts stimulated by PPNLE had an effect on the induction of tube formation, by enhancing VEGF receptor-2 (VEGFR-2) phosphorylation and activation of VEGFR2-mediated downstream pathways such as Protein kinase B (AKT) and extracellular signal-regulated kinase (ERK) in activation of endothelial cells for angiogenesis during wound repair. Moreover, in an in vivo rat model, it has been shown that PPNLE markedly improved skin injury by regulating collagen deposition, re-epithelialization, and neovascularization. These results suggest that PPNLE could be developed as a drug for skin wound healing.

Pilonidal sinus disease (PSD) is a chronic inflammatory condition thought to result from the insertion of external hairs through the epidermis, effectively leading to inflammation and cyst formation. Presenting clinical features comparable to those of PSD, hidradenitis suppurativa (HS) is also characterized as a chronic inflammatory skin disorder associated with hormonal and gene dysregulation, although the exact etiology remains unclear. Given the overlapping clinical features between PSD and HS, this study aimed to evaluate the histologic and immunohistochemical differences between PSD and HS. Using 70 patient tissues and 19 normal skin controls in formalin-fixed paraffin-embedded tissues, protein expressions of cytokeratin 5/6, KLK7, filaggrin, envoplakin, and EPHA2 were analyzed in the epithelium of PSD and HS lesions using immunohistochemistry. Additionally, iron levels, hair shaft presence, and multinucleated macrophage counts were compared, along with disease prevalence across sex and ethnicity. PSD lesions exhibited higher iron levels, and more frequent intralesional hair shafts than HS. The condition was noted more frequently in younger White males while HS was more frequently found in older African American females. The immunohistochemical assays determined that cytokeratin 5/6, KLK7, filaggrin, envoplakin, and EPHA2 increased in lesional skin. The results support the theory that the immune and epithelial response in PSD and HS are similar despite their mechanistically divergent origins.

This study investigated the impact of crocin, a carotenoid component of saffron, on liver damage induced by myocardial infarction (MI) in melatonin deficiency. A synthetic catecholamine called isoproterenol (ISO) was utilised to cause MI-like lesions in rats, simulating the symptoms of heart failure. Using 70 Wistar Albino rats, the following groups were established: (i) control, (ii) sham, (iii) pinealectomy (PNX), (iv) isoproterenol (85 mg/kg), (v) PNX + ISO, (vi) PNX + Crocin (30 days/50 mg/kg), and (vii) PNX + ISO + crocin. To evaluate liver damage, histological, immunohistochemical, and biochemical analyses were performed. MI significantly increased oxidative stress markers such as malondialdehyde and decreased antioxidant levels (glutathione, superoxide dismutase, catalase). Crocin treatment improved oxidative stress markers compared to the untreated groups. Elevated liver enzymes (aspartate aminotransferase, alanine transaminase, alkaline phosphatase) confirmed liver injury in the ISO groups. The levels of these enzymes were not substantially changed by crocin treatment. The liver tissue from the ISO and PNX groups showed moderate-to-severe damage, including inflammation, apoptosis, and hepatocyte degeneration. Crocin treatment reduced these histopathological changes. Crocin reduced the expression of Caspase-3 and increased Ki-67 expression, suggesting its potential to inhibit hepatocyte apoptosis and promote liver regeneration. Crocin treatment showed hepatoprotective effects by reducing oxidative stress, liver enzyme levels, and histopathological damage. More research is required to fully understand the mechanisms of crocin's protective actions and evaluate its clinical applicability.