The Cutting and Floating Method for Paraffin-embedded Tissue for Sectioning

Qian, Cheng; Bai, Yijiang; Zeng, Zhen; Wang, Liao; Luo, Zhen; Wang, Shunqi; Zou, Suqi

doi:10.3791/58288-v

Cited by 4 publications

(3 citation statements)

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“…The cells were treated with different treatments after inoculation and then incubation was continued for 24 h before staining according to the β ‐galactosidase ( β ‐gal) staining kit 47 …”

Section: Methodsmentioning

confidence: 99%

Erxian Decoction‐induced serum exosomes slowed bone marrow mesenchymal stem cell senescence through mitophagy

Huang,

Qian,

Feng

et al. 2023

The Journal of Gene Medicine

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ObjectiveErxian Decoction (EXD) is traditionally employed in the treatment of menopausal syndromes, although its underlying mechanisms remain largely undefined. Given that the senescence of bone marrow mesenchymal stem cells (BMSCs) is intertwined with organismal aging and associated diseases, this study endeavored to elucidate the influence of EXD on aging BMSCs and uncover the mechanisms through which EXD impedes BMSC senescence.MethodsInitially, we probed the anti‐senescent mechanisms of EXD on BMSCs via network pharmacology. We subsequently isolated and identified exosomes from the serum of EXD‐fed rats (EXD‐Exos) and administered these to H2O2‐induced aging BMSC. Assays were conducted to assess BMSC senescence indicators and markers pertinent to mitochondrial autophagy. Treatments with mitophagy inhibitors and activators were then employed to substantiate our findings.ResultsProtein–protein interaction (PPI) network analyses spotlighted AKT1, TP53, TNF, JUN, VEGFA, IL6, CASP3 and EGFR as focal targets. Gene Ontology and Kyoto Encylcopedia of Genes and Genomes pathway analyses underscored oxidative stress, mitophagy and cell proliferation as pivotal processes. Our cellular assays ascertained that EXD‐Exos mitigated H2O2‐induced senescence phenotypes in BMSCs. Moreover, EXD‐Exos ameliorated disrupted mitophagy in BMSCs, as evidenced by enhanced cellular membrane potential and diminished reactive oxygen species levels. Intriguingly, EXD‐Exos also preserved the osteogenic differentiation potential of BMSCs while curtailing their adipogenic propensity.ConclusionOur findings compellingly suggest that EXD counteracts BMSC senescence by fostering mitophagy.

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Section: Methodsmentioning

confidence: 99%

Erxian Decoction‐induced serum exosomes slowed bone marrow mesenchymal stem cell senescence through mitophagy

Huang,

Qian,

Feng

et al. 2023

The Journal of Gene Medicine

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“…The kidney was made into paraffin sections according to conventional methods. Briefly, the kidney tissues were fixed with 10% formaldehyde, dehydrated, and embedded in paraffin [24]. The sections were dehydrated and stained with hematoxylin and eosin (Beyotime, China).…”

Section: Methodsmentioning

confidence: 99%

Identification of Hub Genes Associated with the Development of Acute Kidney Injury by Weighted Gene Co-Expression Network Analysis

Lin¹,

Li²,

Tan³

et al. 2021

Kidney Blood Press Res

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Background: Acute kidney injury (AKI) is a severe clinical syndrome, causing a profound medical and socioeconomic burden worldwide. This study aimed to explore underlying molecular targets related to the progression of AKI. Methods: A public database originated from the NCBI GEO database (serial number: GSE121190) and a well-established and unbiased method of weighted gene co-expression network analysis (WGCNA) to identify hub genes and potential pathways were used. Furthermore, the unbiased hub genes were validated in 2 classic models of AKI in a rodent model: chemically established AKI by cisplatin- and ischemia reperfusion-induced AKI. Results: A total of 17 modules were finally obtained by the unbiased method of WGCNA, where the genes in turquoise module displayed strong correlation with the development of AKI. In addition, the results of gene ontology revealed that the genes in turquoise module were involved in renal injury and renal fibrosis. Thus, the hub genes were further validated by experimental methods and primarily obtained Rplp1 and Lgals1 as key candidate genes related to the progression of AKI by the advantage of quantitative PCR, Western blotting, and in situ tissue fluorescence. Importantly, the expression of Rplp1 and Lgals1 at the protein level showed positive correlation with renal function, including serum Cr and BUN. Conclusions: By the advantage of unbiased bioinformatic method and consequent experimental verification, this study lays the foundation basis for the pathogenesis and therapeutic agent development of AKI.

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“…Brain tissues were fixed in 10% buffered formaldehyde for a week. Histological preparations were performed as previously described (13). Brain tissues were dehydrated through upgrading levels of ethanol solutions and cleared in xylene then embedded in paraffin.…”

Section: Histological Preparation and Proceduresmentioning

confidence: 99%

Effect of Chronic Unpredictable Mild Stress on Hippocampus and Serum Markers in Rats

Yener,

Çolak,

Özbek

et al. 2023

experimed

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Objective: Unpredictable stress is a common factor that we encounter in our daily lives. In this context, we wanted to investigate the morphological effects of a chronic unpredictable mild stress (CUMS) model on rat hippocampus and serum levels of brain derived neurotrophic factor (BDNF) and glial fibrillary acidic protein (GFAP). Materials and Methods:This study was carried out on 16 Wistar albino rats which were divided into control and experiment groups. The CUMS model protocol was applied, and the morphological structures of the hippocampus were examined. The serum GFAP/BDNF levels were measured by ELISA method. Results:The mean BDNF level was 1.65 ± 0.17 ng/mL in the control group and 2.25 ± 0.29 ng/mL in the CUMS group. The mean GFAP level was 3.04 ± 0.45 ng/mL in the control group and 2.96 ± 0.58 ng/mL in the CUMS group. When the polymorph cell layer (PCL) in gyrus dentatus was examined; the mean number of cells in the control group was 20 ± 1.38 for the left PCL and 17 ± 1.35 for the right PCL. In the stress group; the mean number of cells in the polymorph cell layer of gyrus dentatus was 27 ± 2.08 cells for the left side and 23 ± 2.01 for the right side.Conclusions: As a result, morphological changes were observed in the gyrus dentatus region which has an important role in memory formation. The cells in the polymorph cell layer of gyrus dentatus underwent changes under stress.

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The Cutting and Floating Method for Paraffin-embedded Tissue for Sectioning

Cited by 4 publications

References 0 publications

Erxian Decoction‐induced serum exosomes slowed bone marrow mesenchymal stem cell senescence through mitophagy

Erxian Decoction‐induced serum exosomes slowed bone marrow mesenchymal stem cell senescence through mitophagy

Identification of Hub Genes Associated with the Development of Acute Kidney Injury by Weighted Gene Co-Expression Network Analysis

Effect of Chronic Unpredictable Mild Stress on Hippocampus and Serum Markers in Rats

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