Biotechnic & Histochemistry最新文献

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.

Infertility affects 10-20% of sexually active couples and male-related factors contribute to 30-50% of all cases of infertility. The development of three-dimensional in vitro approaches to promote the differentiation of spermatogonial stem cells (SSCs) into functional spermatozoa is an essential step for the treatment of male infertility. Stromal vascular fraction (SVF) cells derived from adipose tissue are known to have a regenerative effect on spermatogenesis and testicular regeneration after testicular damage. The study assessed the effect of SVF cells from epididymal and inguinal adipose tissue on in vitro spermatogenesis. Testicular cells from prepubertal mice were cultured alone as control and co-cultured with SVF cells from epididymal (ESVF) and inguinal (ISVF) adipose tissue in experimental groups by an air-liquid interface system. Spermatogenic progression was evaluated histomorphometrically and immunohistochemically at weeks 1, 3, 4, and 6. ESVF increased formation of tubule-like structures at week 1 and ISVF had a similar effect at week 4. The ISVF group showed higher numbers of ID4(+) (Inhibitor of DNA Binding 4) SSCs than control at all time points. At weeks 3 and 4, the ISVF exhibited increased number of SCP3(+) (Synaptonemal Complex Protein 3) spermatocytes compared to the control group and the ESVF showed a similar increase at week 6. The presence of ACR(+) (Acrosin) spermatids was observed in all groups at week 3. At week 4, the ISVF group had more ACR(+) spermatids than the control and ESVF groups. Our findings demonstrated that SVF cells effectively supported in vitro spermatogenesis. Notably, inguinal-derived SVF cells led to a higher production of ACR(+) spermatids than edidiymal-derived SVF cells. In conclusion, inguinal derived SVF cells can be used as a new co-culture method to preserve the SSC pool and promote in vitro spermatogenesis in infertile patients.

In salivary glands, the concurrent availability of HCO3^- is required for normal mucus release that is guaranteed by the carbonic anhydrases in this area. Due to lack of detailed carbonic anhydrase data in minor salivary glands, we identified various carbonic anhydrases (CA) in the human labial glands of infants in this study. Specifically, the CA isoenzymes II, III, IV, VI, VII, and XII were investigated in secretory and ductal epithelial cells. CA II was detected in serous glandular cells and sporadically in ductal cells, CA III in myoepithelial cells and skeletal muscle cells, CA IV in ductal cells, endothelial cells, erythrocytes, and skeletal muscle. The first immunohistochemical analysis of CA VII in salivary glands resulted in a positive reaction in serous glandular cells and ductal cells, as well as in skeletal muscle and endothelial cells. CA XII was sporadically localized at the basolateral membrane of ductal cells and in serous glandular cells. Mucous endpieces were negative in all carbonic anhydrases tested. Knowledge of carbonic anhydrases distribution in healthy tissues supports their assessment as biomarkers for cancer diagnosis, prevention, and therapy.

Ulcerative colitis (UC) is a chronic inflammatory bowel disease, and endoplasmic reticulum stress (ERS) may contribute to the pathogenesis and progression of UC. Previous research has found that hsa-miR-10a-5p is abnormally expressed in UC, but its molecular mechanisms remain unclear. This study aimed to explore the role of hsa-miR-10a-5p in regulating ERS in UC through RNF186. LPS-induced HT-29 cells and a dextran sulfate sodium (DSS)-induced animal model were utilized to explore the regulatory roles of hsa-miR-10a-5p and RNF186 in ERS in UC. Following modulation of hsa-miR-10a-5p and RNF186 expression, we assessed the expression of ERS-related genes in the UC model using qPCR and immunofluorescence, and evaluated apoptosis with flow cytometry and WB. Furthermore, we conducted DAI scoring, HE staining, and permeability testing in the animal model, and analyzed the inflammatory profile of UC by ELISA to further understand disease progression and the impact of molecular changes on intestinal pathology. Overexpression of hsa-miR-10a-5p in HT-29 cells and animal models promoted cell proliferation, inhibited apoptosis, and mitigated inflammatory factor release and ERS response. Conversely, miR-10a-5p knockdown activated colonic mucosal epithelial damage, reduced cell proliferation, increased apoptosis, aggravated ERS response, and enhanced inflammatory factor expression. This research elucidated that hsa-miR-10a-5p participates in the process of UC development and progression by modulating the ERS response through RNF186. This discovery offered novel insights, enhancing our comprehension of the underlying pathophysiology of UC and provided a theoretical basis for the potential application of miRNAs in UC therapy.

The ability to escape immune surveillance is a hallmark of malignancy. Programmed death ligand 1 (PD-L1) facilitates tumor progression by binding to the immune inhibitory receptor known as programmed cell death protein 1 (PD1) on immune cells, resulting in suppression of the cytotoxic T lymphocyte function. The degree of PD-L1 expression may have a prognostic value in some cancer types, and it may vary according to the genetic makeup and the ethnicity of patients. The expression level of PD-L1 in 63 cases of primary head and neck squamous cell carcinoma (HNSCC) tumor tissues was evaluated using immunohistochemistry (IHC). Also, PD-L1 association with various clinicopathologic characteristics and overall survival was studied. The positive expression rate of PD-L1 in HNSCC was 85.7%, 60.3%, and 52.3% of the total number of cases using combined positive score (CPS) ≥ 1, CPS ≥ 5, and CPS ≥ 20 cutoff values, respectively. Statistical analysis revealed no significant relationship between the expression of PD-L1 protein and clinicopathological features except for tobacco use using a cutoff CPS ≥ 20. The log-rank chi-square results showed that PD-L1 was not a significant factor affecting the 4-year overall survival of HNSCC patients. Also, the overall survival rate was not significantly affected by the patient's age, tumor differentiation, tumor size, and lymphovascular invasion. However, survival curves demonstrated lower overall survival in HNSCC female patients, disease recurrence, and positive perineural invasion. Our findings showed relatively high PDL-1 expression in most HNSCC patients. No significant association was found between PD-L1 protein expression and overall survival.

Studies have indicated that spleen and white pulp atrophy develops within 5 weeks following hyperglycemia onset in streptozotocin-induced diabetic rats. This study aimed to delineate the histopathological alterations in the spleen across two stages of diabetes progression using design-based stereology. Twenty-six rats were categorized into four groups based on condition (normal control [NC] or diabetic model [DM]) and observation period post-induction (5 or 10 weeks): NC5, DM5, NC10, and DM10. Diabetes was induced using streptozotocin-nicotinamide combination. Histological evaluations were performed using standard staining techniques, whereas spleen compartment volumes were quantitatively assessed through point-counting methods on histological sections. Additionally, immunohistochemistry (IHC) and flow cytometry analyses were utilized to determine the distribution and percentages of T and B lymphocytes. Compared to its NC5 control, the DM5 group exhibited inflammatory responses, including polymorphonuclear leukocyte infiltration, but no significant atrophy. DM5 showed a significantly elevated IHC score for T lymphocytes (p < 0.01) and a higher percentage of CXCR5 + B lymphocytes (p < 0.05) compared to NC5, suggesting an active adaptive immune response. In contrast to the NC10 group, the DM10 group displayed significant spleen atrophy (p = 0.005), with marked reductions in total white pulp volume (p = 0.015) and marginal zone volume (p = 0.008). Furthermore, compared to NC10, DM10 exhibited an increased connective tissue volume fraction (p < 0.001). Across all groups, spleen atrophy was directly correlated with reductions in body weight. These findings underscore an initial inflammatory phase characterized by immune cell recruitment in the spleen during early diabetes, subsequently evolving into significant atrophy, reduced white pulp and marginal zone volumes, and an increased connective tissue volume fraction in advanced stages of the disease, all proportional to body weight loss.