Biotechnic & Histochemistry最新文献

Many attempts have been made to simplify tissue fixation and processing prior to hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC) and genomic DNA extraction. The acetone-methyl benzoate-xylene (AMeX) method, is gaining popularity as an alternative to aqueous formaldehyde fixation followed by alcohol dehydration. We have assessed the efficacy of AMeX for the quality of extracted genomic DNA in addition to H&E and IHC in biopsy specimens of human oral mucosal lesions of size not more than 1 cm. H&E staining showed similar results when AMeX method was compared with conventional fixation and processing. Improved IHC staining quality was observed in the sections of AMeX-processed tissue, as were improved quality and quantity of genomic DNA.

We investigated possible protective effects of chlorogenic acid (CGA) against cyclophosphamide (CP) induced hepatic injury in mice. We measured aminotransferase alanine transaminase (ALT) and aspartate transaminase (AST) levels in the serum. We assayed catalase (CAT), superoxide dismutase (SOD), reduced glutathione (GSH), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in hepatic tissue. We assessed expression of nuclear transcription factor 2 (Nrf2) and Kelch sample related protein-1 (keap1) proteins in hepatic tissues using immunohistochemistry. The relative mRNA expression levels of heme oxygenase-1 (HO-1), NADH quinone oxidoreductase 1 (NQO1), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were determined using quantitative real-time polymerase chain reaction (qRT-PCR). Hematoxylin & eosin staining was used to assess liver histopathology. We found that administration of CGA prior to induction of injury by CP decreased serum ALT, AST and MDA expressions in hepatic tissue, while CAT, SOD, GSH and GSH-Px concentrations were increased. We found that hepatocytes of animals administered CGA gradually returned to normal morphology. CGA increased the protein expression of Nrf2 in murine hepatic tissue. Administration of CGA up-regulated mRNA expression levels of HO-1, NQO1, TNF-α and IL-6 in hepatic tissue. CGA exhibited a marked protective effect on CP induced liver injury in mice.

We have investigated anti-obesity effects of the extract of licorice (Glycyrrhiza glabra) root in rats with diet-induced obesity and hyperlipidemia by using histopathological and biochemical methods. Thirty-two Wistar albino rats were divided to four groups of eight: normal control (C), high fat diet (HFD), high fat Diet + Glycyrrhiza glabra (HFD+M), and normal diet with Glycyrrhiza glabra (M). The high fat diet contained 300 g/kg fat (4000 kcal/kg); the daily dosage of Glycyrrhiza glabra extract was 1g/kg body weight by orogastric gavage. Supplementation of Glycyrrhiza glabra extract dramatically reduced increases in body weight caused by the induction of obesity. A hepatoprotective effect of Glycyrrhiza glabra extract was supported by the almost normal histology in the livers of the HFD+M rats, in contrast to the degenerative changes in the HFD rats, which included macrovesicular and microvesicular fat deposits, hydropic degeneration, dilatation of sinusoids and coagulation necrosis of some hepatocytes. Serum levels of alanine transaminase (ALT), aspartic transaminase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), cholesterol (HDL and LDL) and triglycerides, were ameliorated by Glycyrrhiza glabra extract treatment. We conclude that Glycyrrhiza glabra extract given together with HFD could prevent obesity and reduce liver damage in rats.

Berberine (BER) is a naturally occurring alkaloid with a multitude of beneficial effects on human health. Although it is one of the most studied phytochemicals, its curative effect against ovarian damage caused by 5-fluorouracil (5-FU) has not been demonstrated to date. The aim of this study was to investigate the possible protective effect of BER against 5-FU-induced ovotoxicity, focusing on its ability to attenuate oxidative stress, inflammation and apoptosis. The 30 female rats were randomly divided into five groups: Control, BER (2 mg/kg), 5-FU (100 mg/kg), 5-FU+BER (1 mg/kg) and 5-FU+BER (2 mg/kg). The levels of malondialdehyde (MDA), total oxidant status (TOS), total antioxidant status (TAS), superoxide dismutase (SOD), catalase (CAT), 8-hydroxy-2'-deoxyguanosine (8-OHdG), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and caspase-3 were determined using spectrophotometric methods. In addition, ovarian samples were evaluated histopathologically using hematoxylin&eosin staining method. The MDA, TOS, 8-OHdG, IL-6, TNF-α and caspase-3 levels significantly increased by 5-FU administration. Also, we found that 5-FU significantly decreased TAS, SOD and CAT levels. Treatments with BER significantly attenuated the 5-FU-induced ovarian damage via increasing the antioxidant capacity and reducing the oxidative stress, inflammation and apoptosis in a dose-dependent manner. Moreover, the ovoprotective effect of BER was also confirmed by histopathological evaluation. BER may be evaluated as a potential candidate molecule to reduce 5-FU-induced ovarian toxicity.

This study investigated whether abemaciclib (ABE) administration had any adverse effects on ovarian and sex hormones in female rats, and the protective effect of curcumin. Forty female rats were equally divided into the sham control, DMSO, curcumin (CMN), ABE, and ABE+CMN groups. Pharmaceuticals were administered by gavage daily for 28 days. Serum sex hormones were measured in an autoanalyzer operating with a microparticle immunoassay method. In addition, histopathological examination and 8-OHdG expression were performed on the ovarian tissue. Progesterone and testosterone levels were significantly decreased, while estradiol levels were significantly increased, in the ABE group compared to the sham and DMSO groups. In addition, there were significant differences in sex hormone levels in the CMN and/or CMN+ABE groups compared to the ABE group. There was decreased expression of 8-OHdG in the ABE+CMN group compared to the ABE or CMN only groups. This study exhibited that ABE administration can adversely affect functions and histology of the ovarian tissue, but CMN therapy may be protective against the adverse effects on ovarian in ABE-induced rats.

Helicobacter pylori, a curved bacterial rod and causative agent of peptic ulcer and gastric adenocarcinoma, is found as an infectious agent in the stomach of over half of the global population. H. pylori has been identified in oral biofilms and its presence in adenotonsillar tissues has been suggested, with variations in testing methodology both proving and disproving its presence. The current study employed 119 formalin-fixed paraffin-embedded tonsillar tissues from an adult population (n=86) in a major metropolitan city with immunohistochemistry procedures using a monoclonal antibody to determine the incidence of H. pylori in the tonsils. H. pylori was identified in 72.1% of the patients and was associated with Actinomyces spp. in 92.0% of those cases. The high incidence of H. pylori in patients undergoing tonsillectomy suggests that H. pylori may be a contributing factor for tonsillitis and tonsillar hypertrophy. Furthermore, the reservoir for H. pylori in the tonsils may explain why some persons remain refractory to antibiotic treatment for gastric H. pylori.

Hesperetin, a citrus flavonoid, has been a widely studied anticancer agent against many types of cancers, but the exact mechanism of efficacy is still unrevealed. Therefore, this study has attempted to delineate the mechanical aspect of hesperetin's anticancer efficacy against colon cancer using immunoblotting, scanning, and transmission electron microscopic studies. The treatment with hesperetin (25 and 50 µM) has significantly (p < 0.0001) curbed down the proliferation and cell viability of HCT-15 cells in a concentration as well as time dependent manner. Hesperetin was able to achieve this through the induction of caspase-dependent apoptosis. Moreover, hesperetin effectively inhibited phosphorylation of Akt with a parallel increase in PTEN expression thereby inhibiting the PI3K signaling axis, which contributes to the suppression of proliferation. In addition, hesperetin enhanced autophagy through dephosphorylating mTOR, one of the downstream targets of Akt with simultaneous acceleration in Beclin-1 and LC3-II expression levels. Interestingly, hesperetin enhanced the effects of Akt inhibitor LY294002 and mTOR inhibitor rapamycin. This study documented the potential of hesperetin to induce apoptosis through simultaneous acceleration over the autophagic process in colon cancer cells. Thus, hesperetin played a beneficial therapeutic role in preventing colon carcinoma growth by regulating the Akt and mTOR signaling axis.

The present study aimed to investigate the histopathological effects of obstetric gel (OG) on vaginal tissue. In this study, 21 female Wistar albino rats were divided into three groups, comprising seven animals in each group. The first group (group 1) was the control group, the second group (group 2) was the physiological saline (PS) group, and the third group (group 3) was the OG group. In group 1, dilatation was performed using Hegar dilators from Hegar 5 to Hegar 10 without any vaginal application. In group 2, the vagina was washed with a PS-filled applicator. In group 3, the vagina was washed with an OG-filled applicator and Hegar dilators were used to achieve vaginal dilatation. In the group of OG-applied rats, there was an increase in mast cell infiltration, tissue epithelial thickness, and fibrillin-1 levels of the mucosa in the vaginal tissue. The present study is the first to investigate the histopathological effects of OG used for vaginal tissue dilatation in rats. OGs have no early effectiveness in preventing the damage caused by compression of the vaginal wall; however, OGs may have a protective effect against pelvic floor pathologies.