Histology and histopathology Vol.36, nº8 (2021)
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- PublicationOpen AccessPrimary undifferentiated pleomorphic sarcoma in oral-maxillary area: retrospective study and molecular analysis(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) You, Yuanhe; Shi, Rongxin; Dai, Meilu; Du, Zhong; Tian, Zhuowei; Xu, Guisong; Wang, Lizhen; Wang, Lizhen; Wang, Yan'an; Xiao, Mengy. Undifferentiated pleomorphic sarcoma (UPS) in oral-maxillary area is rarely reported. Herein, we aimed to investigate the clinical characteristics, treatment strategies, prognosis, and molecular features of the oral-maxillary UPS. In total, 10 cases with primary oral-maxillary UPS were included. The rapidly progressive UPS can easily develop to an advanced and life-threatening stage, especially concerning the complex anatomical structures and spaces in the oral-maxillary area. The final diagnosis for UPS greatly depended on histological findings and immunohistochemistry staining after the exclusion of all possible differential diagnoses. Retrospectively, the treatment strategies for the included cases still referred to those of oral squamous cell carcinoma (OSCC). Statistically, the median overall survival (OS) for all the included cases was 7.75 months (range: 5-17 months). Comparatively, 3 cases had improved OS (median survival: 17 months, range: 17-18 months) and experienced PR/SD with neoadjuvant chemotherapy (anlotinib). The molecular features were demonstrated by using whole exonic sequencing for 1 included case. Cancer driver gene detection revealed GBP4 as a candidate driver gene for the primary oralmaxillary UPS. Additionally, a missense mutation in gene PIK3CA (p.E545K) was also identified. Our findings could greatly expand the knowledge about primary oral-maxillary UPS, and provide molecular evidences to improve the therapeutic options for primary oral-maxillary UPS.
- PublicationOpen AccessPorcine endometrial 3D co-culture: Morphological changes in 3D endometrium tissues according to hormonal changes(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Kim, Sang-Hwan; Park, Yong-Su; Shin, Da-Hye; Moon, Jeong-Chan; Oh, Min-Gee; Yoon, Jong-TaekCells cultured as monolayers proliferate well, but do not sustain their differentiation characteristics. Previous studies have investigated the interactions between cells and growth factors or cytokines by establishing either in vivo or in vitro three-dimensional (3D) cultures. Using porcine uterine epithelial cells and endometrial cells, the current study was designed to develop a 3D uterine culture system and investigate the response to hormone treatment. Formation of the 3D uterine model was similar to that of uterus from the group supplemented with calcium and magnesium, and the addition of these ions altered the spectrum of basement membrane degrading enzyme expression and activity. In particular, the epithelial cell junctions in the 3D model most closely resembled those of an actual uterus when the medium was supplemented with calcium and magnesium; the intercellular basement membrane structure was also tall under these conditions. The study confirmed that Casp-3 expression was lowest in the P4 (progesterone) treatment group, and this hormone was the most potent stimulus for formation of the endometrial cell layer. Therefore, the addition of calcium and magnesium plays an important role in the formation of a 3D uterine model, and the addition of P4 hormone mimics uterine thickening by stimulating growth of the epithelial cell layer.
- PublicationOpen AccessOsseointegration of a novel 3D porous Ti-6Al-4V implant material – Histomorphometric analysis in rabbits(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Frosch, Stephan; Buchhorn, Gottfried; Krohn, Sebastian; Lehmann, Wolfgang; Frosch, Karl-Heinz; Füzesi, László; Frosch, AlicePorous structure properties are known to conduct initial and long-term stability of titanium alloy implants. This study aims to assess the histomorphometric effect of a 3-D porosity in Ti-6Al-4V implants (PI) on osseointegration in comparison to solid Ti-6Al-4V implants (SI). The PI was produced in a spaceholder method and sintering and has a pore size of mean 400 µm (50 µm to 500 µm) and mimics human trabecular bone. Pairs of PI and equal sized SI as reference were bilaterally implanted at random in the lateral femoral condyle of 16 Chinchilla-Bastard rabbits. The animals were sacrificed after 4 and 12 weeks for histomorphometric analysis. The histomorphometric evaluation confirmed a successful short-term osseohealing (4 weeks) and mid-term osseoremodeling (12 weeks) for both types of implants. The total newly formed bone area was larger for PI than for SI after 4 and 12 weeks, with the intraporous bone area being accountable for the significant difference (p<0.05). A more detailed observation of bone area distribution revealed a bony accumulation in a radius of ±500 µm around the implant surface after remodeling. The boneto-implant contact (BIC) increased significantly (p<0.05) from 4 to 12 weeks (PI 26.23% to 42.68%; SI 28.44% to 47.47%) for both types of implants. Due to different surface properties, however, PI had a significant (p<0.05) larger absolute osseous contact (mm) to the implant circumference compared to the SI (4 weeks: 7.46 mm vs 5.72 mm; 12 weeks: 11.57 mm vs 9.52 mm [PI vs. SI]). The regional influences (trabecular vs. cortical) on bone formation and the intraporous distribution were also presented. Conclusively, the porous structure and surface properties of PI enable a successful and regular osseointegration and enhance the bony fixation compared to solid implants under experimental conditions.
- PublicationOpen AccessMaternal protein restriction changes structural and metabolic gene expression in the skeletal muscle of aging offspring rats(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Valente, Jéssica Silvino; Perez, Érika Stefani; Zanella, Bruna Tereza Thomazini; Gutierrez de Paula, Tassiana; Alcantara dos Santos, Sérgio Alexandre; Oliveira da Silva Duran, Bruno; Carvalho, Robson Francisco; Justulin, Luis Antonio; de Almeida Fantinatti, Bruno Evaristo; Dal-Pai-Silva, MaeliMaternal protein restriction affects postnatal skeletal muscle physiology with impacts that last through senility. To investigate the morphological and molecular characteristics of skeletal muscle in aging rats subjected to maternal protein restriction, we used aged male rats (540 days old) born of dams fed a protein restricted diet (6% protein) during pregnancy and lactation. Using morphological, immunohistochemical and molecular analyses, we evaluated the soleus (SOL) and extensor digitorum longus (EDL) muscles, muscle fiber cross-sectional area (CSA) (n=8), muscle fiber frequency (n=5) and the gene expression (n=8) of the oxidative markers (succinate dehydrogenase-Sdha and citrate synthase-CS) and the glycolytic marker (lactate dehydrogenase-Ldha). Global transcriptome analysis (n=3) was also performed to identify differentially regulated genes, followed by gene expression validation (n=8). The oxidative SOL muscle displayed a decrease in muscle fiber CSA (*p<0.05) and in the expression of oxidative metabolism marker Sdha (***p<0.001), upregulation of the anabolic Igf-1 (**p<0.01), structural Chad (**p<0.01), and Fmod (*p<0.05) genes, and downregulation of the Hspb7 (**p<0.01) gene. The glycolytic EDL muscle exhibited decreased IIA (*p<0.05) and increased IIB (*p<0.05) fiber frequency, and no changes in muscle fiber CSA or in the expression of oxidative metabolism genes. In contrast, the gene expression of Chad (**p<0.01) was upregulated and the Myog (**p<0.01) gene was downregulated. Collectively, our morphological, immunohistochemical and molecular analyses showed that maternal protein restriction induced changes in the expression of metabolic, anabolic, myogenic, and structural genes, mainly in the oxidative SOL muscle, in aged offspring rats
- PublicationOpen AccessHistone demethylase PHF8 promotes cell growth and metastasis of non-small-cell lung cancer through activating Wnt/β-catenin signaling pathway(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Hu, Yan; Mu, Hanshuo; Yang, YingPHD finger protein 8 (PHF8), serving as a histone demethylase, is upregulated in some types of malignant tumors. The role of PHF8 in non-small-cancer lung carcinoma (NSCLC) remains unclear. This study aims to verify the effect of PHF8 in NSCLC and its molecular mechanism. We collected 20 cases of fresh NSCLC and adjacent lung tissues to assess differential expressions of PHF8 by reverse transcriptionquantitative PCR (RT-qPCR). Western blot was employed to examine protein levels of PHF8, Wnt1, βcatenin and epithelial-mesenchymal transition (EMT) related proteins. Chromatin immunoprecipitation assays were executed to confirm the regulatory mechanism of PHF8 and Wnt1. Cell Counting Kit-8 assays and Transwell assays were utilized to identify the effects of PHF8/Wnt1 pathway on cell proliferation, migration and invasion. PHF8 was overexpressed in NSCLC tissues and cells and higher PHF8 expression was correlated with poorer overall survival in NSCLC patients. PHF8 overexpression promoted NSCLC cell proliferation, migration and invasion, while PHF8 knockdown exerted the opposite effect. Mechanistic investigations identified that PHF8 occupied the Wnt1 promoter, leading to a decrease of repressive histone markers H3K9me1, H3K9me2, H3K27me2 and H4K20me1 in the promoter region of the Wnt1 gene, which further promoted the transcription of the Wnt1 gene. PHF8 activated Wnt/βcatenin signaling pathway through promoting Wnt1 expression. Besides, PHF8 altered the EMT of NSCLC through regulating Wnt1 levels. PHF8, acting as an oncogene and prognostic biomarker in NSCLC, stimulated NSCLC to proliferate, metastasis and EMT by activating Wnt/β-catenin signaling.
- PublicationOpen AccessTraumatic brain injury: Estimate of the age of the injury based on neuroinflammation, endothelial activation markers and adhesion molecules(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) dell'Aquila, Massimiliano; Maiese, Aniello; De Matteis, Alessandra; Viola, Rocco Valerio; Arcangeli, Mauro; La Russa, Raffaele; Fineschi, VittorioStudies on traumatic brain injury (TBI) are applicable not only in the clinical context, but also in the forensic field. Over time, the literature has accumulated scientific evidence supporting the use of specific histopathological tests in dating traumatic brain injuries. In primary damage, cell death occurs by necrosis/apoptosis. In secondary injury, the underlying mechanisms are inflammation and ischemia. The inflammatory response of the central nervous system (CNS) follows the common steps of the innate response. In head injury, the blood brain barrier (BBB) undergoes both functional damage and, subsequently, finer structural changes. Scientific evidence has shown modifications of the junctional-endothelial system that favors the extravasation of immunocompetent cells. The histological evaluation of the subdural hematoma, of the cerebral contusions, of the diffuse axonal damage can certainly bring useful elements, with limitations, to the chronological evaluation of the lesions. Many markers have been used to better define the dating of the head injury. Several authors also analyzed the usefulness of secondary damage markers in brain tissue. The progress achieved with immunohistochemistry is significant compared to the use of routine staining. With immunohistochemistry it is possible to identify much narrower and more precise time intervals and, above all, with greater probative reliability. Recently attention has been paid to the modification of structural proteins and miRNAs. Future research is already started and entrusted to multidisciplinary teams that know how to combine their specific skills in search of a reproducible standard of known and sufficient accuracy
- PublicationOpen AccessUpdate on molecular biomarkers for diagnosis and prediction of prognosis and treatment responses in gastric cancer(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Sasahara, Masahiro; Kanda, Mitsuro; Kodera, YasuhiroGastric cancer (GC) is one of the leading causes of cancer-related deaths worldwide, and its high mortality rate is a serious problem in many regions. To improve prognosis, it is necessary to identify novel biomarkers for the early detection of GC, along with its prognosis, risk of metastatic recurrence, and predicted response to chemotherapy, and to develop individualized treatment strategies. Advances in microarray and sequencing techniques have led to the elucidation of cancer-related gene mutations and aberrant expression levels, which have deepened our knowledge of GC. Further searches for sensitive biomarkers are needed to improve the management of patients with GC. In this review article, we update the current knowledge of GC biomarkers, examine recently published literature, and introduce some representative molecules.
- PublicationOpen AccessThe function and regulation mechanism of piRNAs in human cancers(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Wu, Wu; Lu, Bing-feng; Jiang, Ru-qi; Chen, ShuoPiwi-interacting RNAs (piRNAs) are mainly expressed in mammalian germ cells, playing an important role in maintaining germtable line DNA integrity, inhibiting transposon transcription and translation, participating in heterochromatin formation, epigenetic regulation, and germ cell genesis. They combine with P-element induced wimpy testis (PIWI) proteins to form effector complexes known as piRNAinduced silencing complexes (pi-RISC) to regulate the gene silencing pathway. Recent evidence suggests that numerous piRNAs, with tumor-promoting and tumorsuppressing functions in cancer development, are dysregulated in tumor tissues, and are related to clinical prognosis. In the present review, we summarize the current state of knowledge on the function and regulatory mechanisms of piRNAs in the tumorigenesis and progression of cancer, providing evidence for the potential use of piRNAs in the diagnosis and clinical treatment of cancer.
- PublicationOpen AccessThe role and mechanism of PKM2 in the development of LPS-induced acute kidney injury(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Wu, Jiajun; Rong, Shu; Zhou, Jing; Yuan, WeijieA previous study suggested that pyruvate kinase M2 (PKM2) plays a vital role of metabolic reprogramming in the regulation of the innate inflammatory response, while PKM2 is a sensitive biomarker for nephrotoxicity. In this study, we investigated the role and mechanism of PKM2 in development of LPS-induced acute kidney injury. The AKI model of mice was established using LPS. The serum levels of blood urea nitrogen (Bun), creatinine (Cr), and Cystatin C (CysC) were identified using the enzyme-linked immunosorbent assay (ELISA). Hematoxylin and Eosin staining (H&E) was employed to assess pathological changes in kidney tissues of LPSinduced AKI model. Immunohistochemical staining and Western blot analysis were carried out to determine the expression of apoptosis-related factors at protein levels. We found that Bun, CysC, and Cr were significantly increased in the LPS group compared with the control group. The histopathological assay showed model swollen tubular epithelial cells and the presence of vacuolar degeneration in the LPS-induced AKI. In addition, expression levels of PKM2 significantly increased in the LPS group compared with the control group at both protein and mRNA levels (P<0.01). The inhibition of PKM2 by shikonin notably suppressed the expression of HIF-1α and apoptosis-related factors such as BNIP3, Bax, and Caspase-3, while the inhibition of PKM2 by shikonin significantly improved the histopathological symptoms of LPS-induced AKI. This study demonstrated the potential role of PKM2 in LPSinduced AKI and identified PKM2 as a promising therapeutic target in the treatment of AKI.