Histology and histopathology Vol.34, nº7 (2019)
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- PublicationOpen AccessHistopathology of human small intestinal and colonic ischemia-reperfusion: Experiences from human IR-models(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Leenarts, Claire A.J.I.; Grootjans, Joep; Hundscheid, Inca H.; Schellekens, Dirk H.S.M.; Lenaerts, Kaatje; Buurman, Wim A.; Dejong, Cornelis H.C.; Derikx, Joep P.M.Intestinal ischemia-reperfusion (IR) injury is a frequent, but potentially life-threatening condition. Although much has been learned about its pathophysiology from animal IR models, the translation to the human setting is imperative for better understanding of its etiology. This could provide us with new insight into development of early detection and potential new therapeutic strategies. Over the past decade, we have studied the pathophysiology of human small intestinal and colonic ischemia-reperfusion (IR) in newly developed human in vivo IR models. In this review, we give an overview of new insights on the sequelae of human intestinal IR, with particular attention for the differences in histopathology between small intestinal and colonic IR.
- PublicationOpen AccessPrimary extranodal vaginal non-hodgkin lymphoma: Diagnostic pitfalls and therapeutic challenges(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Petrillo, Marco; Fara, Antonella Maria; Fedeli, Maria Antonietta; Fozza, Claudio; Cossu, Antonio; Gulotta, Alessandra; Dessole, Francesco; Piana, Andrea; Capobianco, Giampiero; Tanda, Francesco; Dessole, SalvatoreIntroduction. Primary extranodal nonhodgkin vaginal lymphoma (PeNHVL) represents a rare entity, with few data published until now. We present here a series of patients with PeNHVL, analyzing our data as part of a detailed review of the available literature. Methods. The study included a consecutive series of 6 patients with final diagnosis of PeNHVL admitted at our Institution between January 2000 and December 2017. The systematic review was conducted according to PRISMA guidelines. A literature search of the PubMed, MEDLINE and EMBASE electronic databases was performed using the following terms: ‘vaginal lymphoma’. Relevant data were collected and analyzed for the purposes of this study, reporting results through a narrative approach. Results. In our series discomfort and vaginal pain, refractory to medical treatments represent the symptoms of disease presentation, and the presence of localized/diffused anelastic area in the vaginal wall with tactile sensation of cork emerges as diagnostic sign (Cork Wall sign). The literature revision included 41 studies, with an overall population of 74 patients. The vast majority of women were diagnosed as early stage disease (93.6%) and received chemotherapy (74.6%) with a very high response rate (96%). Death from disease occurred in 5 women (6.7%). Conclusions. Localized or diffused hard-ligneous vaginal areas with Cork Wall sign represent the typical sign of disease presentation. PeNHVL is characterized by a very high sensitivity to chemotherapy and very favourable prognosis; therefore, radical surgery is not indicated. Histotype characterization is crucial to identify those uncommon variants associated with a less favorable clinical outcome.
- PublicationOpen AccessReduction of PGRN increased fibrosis during skin wound healing in mice(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Li, Shan-shan; Zhang, Mei Xiang; Wang, Yue; Wang, Wei; Zhao, Chun Ming; Sun, Xiao Ming; Dong, Guo Kai; Li, Zhou Ru; Yin, Wen Jiang; Zhu, Bo; Cai, Hong XingProgranulin (PGRN) is a multi-functional growth factor known to be involved in regulating of development, cell cycle progression, cell motility, tumorigenesis and angiogenesis. Research has revealed that PGRN is a crucial mediator of skin wound healing. Nonetheless, the role of PGRN in the fibrosis process of cutaneous wound healing has not been identified. In the present study, mice with excisional wounds were treated with si-m-PGRN or physiological saline. We observed the expression of PGRN in intact and post-injury skin by immunohistochemistry. Tissue sections of skin around the wound were performed by hematoxylin and eosin and masson's trichrome staining. After PGRN knockdown by siRNA, the expression of PGRN, collagen I (Col I), small mothers against decapentaplegic homolog 3 (Smad3), phosphorylated Smad3 (P-Smad3), transforming growth factor (TGF)-β1 and TGF-β receptor I (TβRI) were detected by real-time reverse transcription polymerase chain reaction (RT-qPCR) or Western blot. PGRN mRNA and protein expressions were increased after insult and remained above that of intact skin through day 20. Down-regulation of PGRN augmented fibrosis area, skin thickness and the expression of Col I. In addition, reduction of PGRN considerably increased the expression of TGF-β1, TβRI, Smad3 and P-Smad3. These results indicate that PGRN knockdown enhances the fibrosis degree, probably via the TGF-β/Smad signaling pathway.
- PublicationOpen AccessMorphological rearrangement of the cortical region, in aging ovaries(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Díaz Hernández, Verónica; Caldelas, Ivette; Montaño, Luis M.; Merchant Larios, HoracioThe ovary is a structurally dynamic organ that alters with age. Modifications in the paracrine status influence the capacity of aging oocytes to develop normal embryos. Despite the importance of understanding the cellular and molecular mechanism involved in the process of ovarian aging, histological changes remain poorly understood. Correlating the process of folliculogenesis and somatic cell function during ovarian aging is essential to explain the reproductive decline of aged mammalian species, including humans. Here, we performed a morphological and immunohistological study on the ovaries of chinchilla rabbits that varied in age from one to 34-months. The spatiotemporal expression of the cholesterol side-chain cleavage cytochrome P450scc (CYP11A) and the smooth muscle actin (SMA) were analyzed. A significant histological rearrangement of immunodetected cells in theca interna, theca externa and the interstitial tissue around the follicles occurred. The expression of CYP11A1 decreased considerably in antral follicles of aging ovaries. Moreover, we found that the secondary interstitial gland developed extensively, and a remarkable rearrangement of the surface epithelium occurred in aging ovaries. In contrast to ovaries during the reproductive period, the immunohistological changes demonstrate that the interstitial gland became the most abundant tissue during the aging of ovaries. Thus, the current study provides new data for understanding the alteration of somatic cell function in elderly ovaries and how this affects their declined fertility.
- PublicationOpen AccessCharacterization of the early pathology of cochlear stereocilia in four inbred mouse strains with progressive hearing loss(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Liu, Xiang; Xie, Yi; Huang, Shanshan; Xu, Ang; Zhao, Mengmeng; Kang, Xiaoxia; Yan, Aiwei; Li, Ping; Jin, Changzhu; Han, FengchanObjective. Inbred strains of mice offer promising models for understanding the genetic basis of age-related hearing loss (AHL). NOD/LtJ, A/J, DBA/2J and C57BL/6J mice are classical models of age-related hearing loss and exhibit early onset of pathology of AHL. This study was carried out to characterize the early pathology of cochlear stereocilia in the four mouse strains with age-related hearing loss. Methods. The structural features of stereocilia in NOD/LtJ, A/J, DBA/2J and C57BL/6J mice were observed by scanning electron microscopy (SEM) at age of 2, 4, 6 or 8, and 10 or 12 weeks. Meanwhile, auditoryevoked brainstem response (ABR) and distortion product otoacoustic emission (DPOAE) amplitudes of the mice were measured at various intervals (3, 4, 6, 8, 10 and 12 weeks of age). Results. The ABR thresholds in NOD/LtJ, A/J and DBA/2J mice increased with age from 3 to 12 weeks. DPOAE amplitudes in NOD/LtJ, A/J, DBA/2J mice were very low at 4 weeks and became negative at 8 weeks at f2 frequency of 17 672 Hz. In addition to the progressive hearing loss, the four mouse strains displayed early onset (at 2 weeks of age) and progressive degeneration of stereocilia in hair cells. Conclusion. Early degeneration of stereocilia contributes to the functional impairment of hair cells and hearing loss in NOD/LtJ, A/J, DBA/2J and C57BL/6J mice.
- PublicationOpen AccessSafety of isotretinoin treatment as measured by liver parameters(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Thomazini, B.F.; Lamas, C.A.; Dolder, M.A.H.Isotretinoin is an analogue of vitamin A and by suppressing the sebaceous glands it is often prescribed in cases of severe acne treatment. The treatment for the average patient is carried out during two to ten months. This study was designed to investigate liver structure, hepatic enzyme levels and the stress oxidative parameter after isotretinoin treatment during a similar period and using the dosages of 1 mg/kg and another one of 10 mg/kg in young male Wistar rats. We have analyzed the blood serum biochemical levels to determine hepatic function and lipid peroxidation, hepatic tissue levels of hepatic enzymes, histology and ultrastructure. The groups receiving 1 mg/kg were not altered after treatment. Their ultrastructure showed a metabolically more active organ after treatment with 10 mg/kg, in which there was an increase in the area occupied by mitochondria and rough reticulum in electron transmission images. The group that received 10 mg/kg also showed increased alkaline phosphatase, decreased high density lipoprotein and low density lipoprotein. The changes observed with the 10 mg/kg dose were not conclusive for liver damage, because of the lack of histological structural modifications and the few biochemical alterations. The 1 mg/kg dose showed a liver responding to some stimuli but without profound alterations. So, we confirm that the proposed protocol with 1 mg/kg or 10 mg/kg isotretinoin did not cause important biochemical and histological disfunctions for male Wistar rat livers.
- PublicationOpen AccessSynergism of imatinib, vatalanib and everolimus in the prevention of chronic lung allograft rejection after lung transplantation (LTx) in rats(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Keil, Laura; Schaub, Anna Lena; Hirt, Stephan W.; Schmid, Christof; Lehle, Karla; Suesskind Schwendi, Marietta vonChronic lung allograft dysfunction (CLAD) still remains a major drawback in the outcome following lung transplantation (LTx). New therapeutic strategies are warranted. Growth factors and their receptors like platelet-derived growth factor-receptor (PDGFR) and vascular endothelial growth factor-receptor (VEGFR), may play a crucial role in the development of CLAD, especially bronchiolitis obliterans (BO) and vasculopathy. In this study, we used an orthotopic left lung transplantation model from Fischer (F344) to Wystar Kyoto (WKY) rats to investigate the effect of the receptor tyrosine kinase inhibitor (RTKI) vatalanib alone, the dual combination of the RTKIs vatalanib and imatinib and a triple therapy consisting of vatalanib, imatinib and the mammalian target of rapamycin inhibitor (mTORI) everolimus on the development of CLAD after LTx in rats. With this trial we demonstrated that monotherapy with vatalanib attenuated mild and severe chronic vascular rejection, whereas dual therapy (vatalanib and imatinib) after LTx also showed a significant reduction of chronic bronchiolar rejection and interstitial fibrosis. By adding everolimus, the effect of vatalanib and imatinib could additionally be increased. In conclusion, the combination of mTORI and RTKIs might be a possible strategy in the prevention of CLAD and BO.
- PublicationOpen AccessPolymorphous adenocarcinoma of the salivary glands: an overview of immunohistochemical features and insights on molecular pathology(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Tasoulas, Jason; Tsourouflis, Gerasimos; Klijanienko, Jerzy; Theocharis, StamatiosPolymorphous adenocarcinoma (PAC), represents a common minor salivary gland tumor (SGT) characterized by local growth, low metastatic potential and non-aggressive biologic behavior. Due to the clinical aggressiveness noted in a subset of such tumors, the former term polymorphous low-grade adenocarcinoma (PLGA) was recently revised. PAC’s clinical features and histological diversity result in clinical overlap of this entity with several other SGTs including mainly adenoid cystic carcinoma (AdCC). Differential diagnosis among the entities is crucial, in terms of tumor management and patients’ prognosis. The aim of the present review is to summarize the histological, cytological, immunohistochemical and molecular features of PAC. Histopathological examination is usually adequate for PAC differential diagnosis from other SGTs, except of AdCC. Several immunohistochemical markers including c-Kit, S-100/ MG, Mcm-2 and Integrin β-1, -3, -4, are reported to be useful diagnostic aids in borderline cases. Limitations in sample numbers and study methodology issues of the immunohistochemical PAC studies complicate the identification and selection of appropriate markers useful in the differential diagnosis. Additionally, molecular analyses of PAC specimens indicate that the PAC spectrum phenotypes result from different genotypes (protein kinase D positive; PRKD(+) and PRKD(-) tumors). PAC pathogenesis remains to be determined in each particular genotype while the convergence issue should be addressed in future studies.
- PublicationOpen AccessHydroxy group requirement for halofuginone-dependent inhibition of muscle fibrosis and improvement of histopathology in the mdx mouse model for Duchenne muscular dystrophy(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Wellner, Gili; Mordechay, Sharon; Evans, Paul; Genin, Olga; Pines, Mark; Halevy, OrnaIn Duchenne muscular dystrophy (DMD), the progressive loss of muscle and its ability to function is associated with significant fibrosis, representing the major disease complication in patients. Halofuginone, a halogenated analog of the naturally occurring febrifugine, has been shown to prevent fibrosis in various animal models, including those of muscular dystrophies. Here, two optically active enantiomers of deoxyhalofuginone—a halofuginone analogue in which the hydroxy group in position 3 was removed from the piperidinyl entity—were evaluated with respect to their effect on muscle histopathology in mdx mice. Male mdx mice were treated with either deoxyhalofuginone (as single enantiomers or in racemic form), or halofuginone, for 10 weeks, starting at the age of 4 weeks. Halofuginone caused a significant reduction in total collagen content, degenerative areas, as well as in utrophin and phosphorylated-Smad3 levels in the mdx diaphragms. However, neither the deoxyhalofuginone enantiomers, nor its racemic form had any effect on these parameters. A positive effect of the deoxyhalofuginone (+)-enantiomer was observed on myofiber diameters; however, it was lesser than that of halofuginone. It is concluded that the hydroxy group plays a key role in halofuginone’s effects related to fibrosis in DMD, and points towards the transforming growth factor β/Smad3 signaling pathway being involved in this inhibition. Elucidation of the structure–function relationship of halofuginone, in relation to inhibiting fibrosis in muscular dystrophies, is of the utmost importance for creating the next generation of anti-fibrotic therapies that will be more efficacious and less toxic, hence improving life quality of patients.
- PublicationOpen AccessHigh expression of GPNMB predicts poor prognosis in head and neck squamous cell carcinoma(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Li, Hao; Xiao, Yao; Wu, Cong Cong; Yang, Lei Lei; Cao, Ling Yun; Chen, De Run; Zhou, Jun Jie; Zhang, Wen Feng; Sun, Zhi JunGlycoprotein non-metastatic protein B (GPNMB) is a transmembrane glycoprotein that is highly expressed in several malignancies compared with its expression in matched healthy tissues. The aim of this study was to investigate the clinical characteristics and prognostic value of GPNMB expression in tumor tissue derived from a cohort of patients with head and neck squamous cell carcinoma (HNSCC). GPNMB expression in human HNSCC, oral dysplasia and normal mucosal tissue was evaluated by immunohistochemistry (IHC). The correlations of GPNMB expression with the clinical characteristics of HNSCC were assessed by oneway ANOVA and t test analyses. Survival data were analyzed using Kaplan-Meier analysis and the Cox proportional hazards model. GPNMB was highly expressed in HNSCC tissue compared with dysplasia and normal mucosal tissue. Additionally, a high level of GPNMB expression in HNSCC was associated with poor prognosis (P<0.01). In the analysis of tumor-nodemetastasis (TNM) staging, a high GPNMB expression level in HNSCC tissue, as well as metastatic lymph node tissue, correlated with an advanced N stage. In conclusion, GPNMB was overexpressed in human HNSCC tissue and predicted poor prognosis in human HNSCC tissue. In addition, GPNMB expression was closely correlated with N stage in patients with HNSCC.
- PublicationOpen AccessThe distribution and time-dependent expression of HIPK2 during the repair of contused skeletal muscle in mice(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Zhang, Miao; Zhang, Meng Zhou; Wen, Shu Heng; Sun, Ying Fu; Jiang, Peng Hao; Wang, Lin Lin; Zhao, Rui; Wang, Chang Liang; Jiang, Shu Kun; Guan, Da WeiHIPK2 is an evolutionarily conserved serine/threonine kinase and is considered a co-regulator of an increasing number of transcription factors modulating a variety of cellular processes, including inflammation, proliferation and fibrosis. Skeletal muscle injuries repair is an overlapping event between inflammation and tissue repair. There are no reports about HIPK2 expression in skeletal muscles after trauma. A foundational study on distribution and time-dependent expression of HIPK2 was performed by immunohistochemical staining, Western blotting and quantitative real-time PCR, which is expected to obtain a preliminary insight into the functions of HIPK2 during the repair of contused skeletal muscle in mice. An animal model of skeletal muscle contusion was established in 50 C57B6/L male mice. Samples were taken at 1, 3, 5, 7, 9, 14, 17, 21 and 28 days after contusion, respectively (5 mice at each posttraumatic interval). 5 mice were employed as control. No HIPK2- positive staining was detected in uninjured skeletal muscle. Intensive immunoreactivties of HIPK2 were observed in polymorphonuclear cells, round-shaped mononuclear cells, regenerated multinucleated myotubes and spindle-shaped fibroblastic cells in the contused tissue. The HIPK2-positive cells were identified as neutrophils, macrophages and myofibroblasts by double immunofluorescent procedure. HIPK2 protein and mRNA expression were remarkably up-regulated after contusion by Western blotting and qPCR analysis. The results demonstrated that the expression of HIPK2 is distributed in certain cell types and is time-dependently expressed in skeletal muscle after contusion, which suggested that HIPK2 may participate in the whole process of skeletal muscle wound healing, including inflammatory response, muscle regeneration and fibrogenesis.