Biotechnic & Histochemistry最新文献

Renal ischemia-reperfusion (Isc-Rep) injury commonly occurs during kidney transplantation, renal vascular surgery, or traumatic hemorrhagic shock, and is marked by excessive oxidative stress and inflammation. We aimed to examine the antioxidant properties of the melatonin-curcumin combination in an experimental renal Isc-Rep model in rats. Thirty-five rats were allocated into five distinct experimental groups. While the rats in the Control (C) group did not undergo any surgical procedure, each of the rats in the remaining groups underwent 45 minutes of renal ischemia followed by 2 h of reperfusion. In all experimental groups, except the Isc-Rep group, the rats were administered an active compound immediately prior to reperfusion, namely melatonin (MEL, 20 mg/kg), curcumin (CUR, 200 mg/kg), or a combination of MEL-CUR. In all groups, histological evaluation was carried out, while apoptotic cell analyses were similarly performed by means of the TUNEL method. In addition, oxidative stress parameters (TOS, TAS, OSI), inflammatory cytokines (TNF-α, IL-6, and IL-1β), superoxide-dismutase (SOD), and malondialdehyde (MDA) activity levels were also examined. Histological evaluation demonstrated that renal Isc-Rep-induced cellular damage, tubular dilatation, and inflammation were significantly reduced in the MEL-CUR groups in comparison with the Isc-Rep group (p < 0.005). Furthermore, statistically significant differences were observed among the groups in TOS, TAS, OSI, TNF-α, IL-6, and IL-1β levels (p < 0.005). The co-administration of melatonin and curcumin resulted in greater protection against ischemia-reperfusion-induced renal damage compared with either treatment alone. Moreover, the co-administration of these antioxidant agents demonstrates greater therapeutic efficacy compared to their individua.

In the present study, an in-situ hybridization (ISH) assay was performed on cell lines and paraffin-embedded tissue blocks infected with infectious pancreatic necrosis (IPN) virus in fish. The molecular diagnosis by ISH was established through four main steps: (1) generation of DIG (Digoxigenin)-labeled probes, (2) production of anti-DIG antibodies and their conjugation with alkaline phosphatase, (3) preparation of ISH reagents, including pre-hybridization and hybridization solutions, and (4) application of the ISH assay to cell culture and paraffin tissue sections. Protease concentrations, hybridization solutions, and assay modifications were optimized separately for cell lines and paraffin sections. Ultimately, ISH assays using DIG-labeled probes targeting the VP2 and VP3 gene regions of IPN virus were successfully performed in virus-infected cell lines and paraffin-embedded tissues.

Parkinson's disease (PD) is a progressive neurodegenerative disorder associated with aging. It is characterized by both motor and non-motor symptoms, including cognitive impairment, mood disturbances, sleep disruption, sexual dysfunction, and weight loss. These symptoms may precede a clinical diagnosis. Sex-related differences in PD, influenced by estrogen's neuroprotective effects and testosterone deficiency, contribute to a higher prevalence and severity of the disease in males. Male rats received daily subcutaneous injections of rotenone (2 mg/kg) for 20 days to establish a PD model, which enabled the investigation of central dopaminergic degeneration and peripheral testicular effects. Tyrosine hydroxylase (TH) immunoreactivity in the substantia nigra pars compacta (SNpc) was assessed alongside the testicular expression of GPR30, GPR120, and GPR125 in the testes using immunohistochemical and immunofluorescence techniques. Rotenone-treated animals exhibited significant weight loss (z = -4.292, p < 0.001), reduced TH expression in the SNpc, and hallmark PD pathology including Lewy body-like inclusions, Lewy neurites, and halo-like structures around dopaminergic neuron nuclei. Testicular analysis revealed a time-dependent decline in GPR protein expression. GPR30 was found in spermatids and interstitial cells, GPR120 was found predominantly in Leydig cells, and GPR125 was found in interstitial tissue, Sertoli cells, and spermatocytes. These findings suggest that the loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc) may affect testicular function via testosterone-dependent regulation of GPR30, GPR120, and GPR125 expression. This highlights the systemic nature of PD and its potential impact on male reproductive health. Alterations in testicular G Protein-Coupled Receptors (GPCRs) may serve as peripheral biomarkers of disease progression, emphasizing the importance of considering endocrine and reproductive dysfunction in both experimental models and the clinical management of PD.

We examined the clinicopathologic features and immunohistochemical expression of commonly tested biomarkers relevant to targeted therapy in postradiation angiosarcoma (PRAS) of the breast, aiming to better understand tumor biology and provide baseline data on tumor biomarkers to inform future therapeutic strategies for PRAS. Clinicopathologic features, outcomes, and the immunohistochemical expression of tumor biomarkers, including human epidermal growth factor receptor 2 (HER2), programmed death-ligand 1 (PD-L1), and DNA mismatch repair proteins (MMR), were analyzed in breast PRAS specimens diagnosed from 2000 to 2024. A total of 24 breast PRAS specimens from 18 patients were included in the study. All PRAS patients were female, with a median age of 72 years at the time of diagnosis. High-grade tumors were observed in 47%, 43%, and 65% of specimens according to Fédération Nationale des Centers de Lutte Contre le Cancer (FNCLCC), Donnell-Rosen, and Kuba scoring methods, respectively. Surgical intervention, adjuvant chemotherapy, and radiotherapy were administered to 100%, 29%, and 23% of patients, respectively. Five patients (29%) died from the disease during a median follow-up period of 37 months. Exploratory analysis showed there was no significant association between histologic grade and overall survival, regardless of the grading system used. All PRASs were MMR-proficient with a low number of tumor-infiltrating lymphocytes (TILs) and exhibited an absence of HER2 protein expression. PD-L1 was positive in seven (32%) specimens and positively correlated with stromal TILs. Our findings suggest that breast PRAS are genomically stable and poorly immunogenic, and are likely ineligible for any HER2-targeted therapies based on current approval criteria. A subset of specimens demonstrated PD-L1 positivity, underscoring the need for further investigation of immunotherapy with anti-PD1/PD-L1 immune checkpoint inhibitors as a potential treatment option.

Pulmonary fibrosis (PF) is a prevalent adverse effect associated with bleomycin (Bleo) administration. Despite extensive research efforts, there is no effective treatment currently available for PF. The current study investigates the antifibrotic and antioxidant properties of tannic acid (TA), a natural polyphenol known for its potent antioxidant and anti-inflammatory activities, in the context of Bleo-induced PF in rats. Fifty Wistar rats were randomly assigned to five distinct groups for the study. Group 1 served as the control (normal saline). Group 2 received a single dose of Bleo (7.5 U/kg via intratracheal injection) on the 7th day. Groups 3, 4, and 5 were treated with 100, 200, and 400 mg/kg of TA, respectively, administered 7 days prior to and 21 days following Bleo administration. The rats were euthanized 24 hours after the last administration. The protective effects of TA were assessed through histopathological and biochemical analyses, which included measurements of lung index, hydroxyproline (HP), nitric oxide (NO), malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT). Additionally, the influence of TA on tumor necrosis factor-α (TNF-α) levels was evaluated. The results indicated that TA significantly reduced the Bleo-induced increase in lung index, MDA, NO, HP, and TNF-α levels. Furthermore, TA enhanced the activities of CAT, SOD, and GPx, as well as the content of GSH. Additionally, TA mitigated the infiltration of fibroblasts and inflammatory cells and prevented alveolar thickening induced by Bleo. The findings demonstrated that TA exhibits a protective effect against PF induced by Bleo.

Oral submucous fibrosis (OSMF) is a ubiquitous fatal fibrotic mucosal disease with multifactorial etiology and complex pathogenesis. The role of mast cells in the pathophysiology of OSMF remains uncharted territory owing to a dearth of studies. Thus, the present systematic review and meta-analysis aimed to unentangle the mysteric role of mast cells in the pathogenesis, progression of fibrosis and malignant transformation of OSMF. Using various databases, full-text articles that investigated mast cell concentrations in OSMF were entailed for review. A modified Newcastle-Ottawa scale was employed to evaluate the risk of bias in all articles and Review Manager was utilized for meta-analysis. Twenty and fourteen qualified articles, respectively, were included for qualitative and quantitative data synthesis. Progressive amplification of mast cell density is linked with fibrosis-induced malignant transformation of OSMF. The fixed-effect model also confirmed that significantly upregulated mast cell counts have a decreased risk of association with control as well as a significantly increased risk of being associated with early-stage fibrosis and malignant transformation of OSMF. This review authenticates the mechanistic effects of mast cells in the pathogenesis, chronicity, progression of fibrosis and malignant transformation of OSMF.

Diabetes mellitus is a prevalent chronic disease, with diabetic nephropathy being a significant complication that causes structural alterations in the kidneys. The purpose of this research was to examine the effects of nano-chitosan (NCh) on the kidneys of rats with diabetes induced by streptozotocin (STZ). NCh was prepared and characterized using scanning electron microscopy (SEM), energy dispersive x-ray spectroscopy technique (EDX) and zeta potential. A total of 40 male Sprague Dawley albino rats (225 ± 25 grams, 2.5-3 months old) were involved and divided into 4 equal groups, each with 10 rats: Control (Ctrl), non-diabetic-administered with 0.5 mg/kg B.W NCh (NCh), diabetic induced by STZ intraperitoneally (Diabetic), and diabetic treated with NCh at 0.5 mg/kg B.W (Diabetic-NCh). Seven days post-diabetic induction, NCh was inoculated orally for 21 days once a day. Body weight change and kidney function tests (creatinine and urea), as well as renal histopathological, histochemical (collagen fibers), immunohistochemical; alpha smooth muscle actin (αSMA) and transforming growth factor-beta1 (TGF-β1), and ultrastructural studies were involved. The findings indicated that diabetic-NCh treated rats exhibited improvements in body weight and kidney function tests including creatinine and urea, compared to the diabetic rats. NCh treatment enhanced renal tissue architecture and reduced collagen fiber expression. Immunohistochemical analysis showed decreased concentrations of αSMA and TGF-β1 in the Diabetic-NCh group. Ultrastructural studies confirmed the improvement in kidney tissue structure. In conclusion, oral administration of nano-chitosan demonstrated a potential therapeutic effect on kidneys in diabetic rats, suggesting its promise as a treatment strategy for diabetic nephropathy.

Varicocele is one of the most important disorders causing infertility in men, and oxidative stress is one of the most important factors affecting testicular parenchyma damage caused by varicocele. This study explored the effect of anthocyanins on varicocele-induced testis injury in adult Wistar rats by focusing on regulating oxidative stress, Bax and Bcl-2 genes, and protein related to cell death. Rats (n = 32) were divided into four groups: Control (Sham), varicocele, varicocele + anthocyanin, and anthocyanin alone. At the end of the study (week 8), the animals were sacrificed, and H&E staining was used for testicular histopathology. The IHC method was used for the detection of Bax and Bcl-2 protein expression, and TUNEL assays were used to analyze testicular Apoptosis. Additionally, serum levels of oxidative stress markers - MDA, SOD, and GPx - were assessed by ELISA, and RT-qPCR analyzed the mRNA expression of Bax and Bcl-2. Histological analysis revealed notable improvements in Johnsen's score, epithelial thickness, and seminiferous tubule diameter in the varicocele + anthocyanin group relative to the varicocele-only group (p < 0.005). Protein and mRNA expression of Bax significantly increased in the varicocele group (p < 0.005), while treatment with anthocyanin enhanced Bcl-2 expression (p < 0.005). Furthermore, the rate of apoptotic positive germ cells decreased when the rats received anthocyanin. Moreover, anthocyanin increased serum levels of GPx and SOD while decreasing MDA levels in the treatment group compared to rats with varicocele (p < 0.005). These outcomes suggest that anthocyanin may moderate testicular injury from varicocele, primarily through its antioxidative properties.

It has been suggested that adipokines may modulate plasma lipid levels, and β3-adrenergic receptor (β3-AR) gene expressions may affect adipokine levels and play critical roles in lipid metabolism. This study aims to determine predictive biomarkers for preterm birth (PTB) by evaluating serum complement 1q (C1q)/tumor necrosis factor (TNF)-related protein (CTRP) levels, lipid profiles, gene expressions, and placental pathological changes in women experiencing PTB. A total of 80 pregnant women, 40 preterm and 40 term, who applied to the Department of Obstetrics and Gynecology, Faculty of Medicine, Inonu University, were included in the study. Blood and placenta samples were taken from all participants. Serum CTRP, high-density lipoprotein (HDL), low-density lipoprotein (LDL), total cholesterol (TC), and triglyceride (TG) levels were measured; β3-AR gene expression and detection rate of the β3-AR rs4994 (Trp64Arg) amplicon were evaluated. Placental tissues were examined histopathologically for perivillous/intervillous fibrin deposition and hydropic degeneration. ROC analysis was used to determine predictive biomarkers for PTB. In the PTB group, compared to the control group, β3-AR gene expression levels and serum CTRP3 levels were significantly decreased, while the detection rate of the Trp64Arg amplicon and serum CTRP4 levels were significantly increased. In addition, LDL levels increased significantly (p = 0.046), TC levels decreased (p = 0.045). According to ROC analysis, LDL (p = 0.039), TC (p = 0.034), CTRP3 (p = 0.019), and CTRP4 (p = 0.033) levels were determined as significant predictive biomarkers for PTB. Histopathological examination revealed increased perivillous and intervillous fibrin deposition and marked hydropic degeneration in the PTB group. Changes in CTRP levels, lipid profile disorders, and a decrease in β3-AR signaling pathways were found to be associated with PTB. LDL, TC, CTRP3, and CTRP4 levels can be evaluated as potential biomarkers that can be used in the early diagnosis and management of PTB.

Acute kidney injury (AKI) is a clinical syndrome that can be caused by a variety of factors, leading to rapid decline of kidney function and increased morbidity and mortality, whilst also exerting significant economic burden on the affected patient. Salvia miltiorrhiza is a highly valued plant in traditional Chinese medicine (TCM). Tanshinone IIA (Tan IIA) is an important active compound that can be extracted from salvia miltiorrhiza, which has reported anti-inflammatory effects. The objective of the present investigation was to explore the potential effects of Tan IIA on folic acid-induced AKI and elucidate its underlying mechanism. A comprehensive analysis was conducted utilizing the TCM Systematic Pharmacology Database and Analytical Platform database to screen for chemical components and their corresponding targets. Subsequently, by using network pharmacology techniques and Cytoscapes 3.7.2 software, a protein-protein interaction (PPI) network was constructed and analyzed. Through Venn diagram analysis of the DeGeNET, OMIM, PharmGKB, and GeneCards databases using the key word "acute kidney injury," a total of 76 overlapping targets were obtained. Building upon this, a compound-target gene network was constructed and analyzed by Cytoscapes 3.7.2 software, revealing TP53, STAT3, CASP3, VEGFA, and JUN to be pivotal therapeutic targets. Subsequently, an AKI mouse model was established to investigate the renal effects of Tan IIA. By immunohistochemistry, Western blot results showed the Tan IIA ameliorated kidney function by alleviating inflammation, mitigating necrosis of renal tubular cells, promoting their proliferation and attenuating kidney injury. These beneficial effects were found to be achieved by inhibiting the PI3K/AKT signaling pathway and inhibiting the expression of TP53 by Western blot. In conclusion, TP53may be a potential target for folic acid-induced AKI, whilst Tan IIA exerts its renoprotective effects and improves renal function by PI3K/AKT signaling pathways.