Zeitschrift für Zytologie und Histologie

Introduction: The purpose is analyze the biological and cellular regeneration of non-repairable lesions of peripheral nerve by muscle graft enhanced with growth factors.
Methods: Controlled clinical experimental trial, level I evidence. Critical peripheral nerve injuries repair through acellular muscle graft alone versus its enrichment with IGF-1 (10 mg/ml mecasermin). Clinical and functional control for 3 months. Animals were sacrificed at 90-100 days, obtaining samples for macro and microscopy with blue toluidine staining, hematoxylin-eosin and Masson trichrome.
Results: Regenerating axons can pass through a defect length of 15 mm with the addition of an acellular skeletal muscle graft and repopulate the distal nerve. Basal lamina tubes, acting as matrices for the axonal growth, appears that sensory recovery exceeds the rate of motor nerve fibers growth. Finding a number of regenerating axons greater than healthy sciatic, ensures the functional connection of distal axons to their target innervation. In contrast, grafts myelinated fibers are smaller than those of healthy sciatic nerve.
Discussion: Acellular muscle fibers basal lamina define tubes that could support axonal regeneration, acting as matrices for the growth of axons, when grafted into a nerve defect. Stereologic studies biological sections to quantify the increase in the total number of axons, the density thereof and the thickness of the myelin from microscopic examination of histological slices of animals treated with the growth factor IGF-1.

Lung cancer closely resembling Extraskeletal Myxoid Chondrosarcoma (EMC), called “primary pulmonary myxoid sarcoma (PPMS),” is an extremely rare tumor. This report is presented here of a unique case showing EMC-like changes only in metastases from conventional lung adenocarcinoma in an 81-year-old man. After lung tumor resection and chemo-radiotherapy, he underwent the resection of right adrenal metastasis composed of carcinomatous and EMC-like components. There were multifocal transitions between these components. He was treated with additional chemo-radiotherapy for newly developed liver and brain metastases, but he died 3.3 years after the initial surgery. Autopsy revealed EMC-like tumors, with or without minute adenocarcinomatous components, extensively involving widespread organs. Reverse transcription-polymerase chain reaction did not detect EWSR1- CREB1 fusion in the primary tumor or metastases. These features indicated that the current tumor was different from PPMS characterized by EWSR1-CREB1 fusion. We concluded that such EMC-like features represent morphological changes of metastatic lung adenocarcinoma. We believe that this possible occurrence should be recognized for accurate diagnosis of metastatic lung carcinoma.

Cytological specimens are used only once for staining, and cannot be reused after preservation in paraffin blocks. If a technique is developed allowing restaining of precious/rare specimens or those for education, which have been discolored due to long-term preservation, to restore their former states, it will be extremely useful for the reuse of specimens. We performed this study to determine whether discolored specimens can be restored to their former condition by applying an optical clearing agent. For restaining, 131 Papanicolaou (Pap) specimens prepared from 13 to more than 30 years ago were used. Using an optical clearing agent (SCALEVIEW-A2), Pap restaining, endurance tests of the optical clearing agent by repeated use, and immunostaining were performed. After treatment with the optical clearing agent for5-10 minutes, discolored cytological specimens could be stably restained just as the initial Pap specimens. As a result of the immunostaining of specimens treated using this method, both the nucleus (Ki-67) and cell membrane (MOC31) were positively stained. The optical clearing agent did not deteriorate when used repeatedly ten times. Discolored precious cytological specimens could be restored using a optical clearing agent. Specimens processed using this method could also be immunostained, and so this method is extremely useful

Demilunes have historically been considered to be integral components of the secretory tree of mixed salivary glands. Based on preservation of rat sublingual glands by rapid freezing and freeze-substitution fixation, Yamashina et al. have advanced the notion that serous demilunes are artifactual products of chemical fixation, that swollen mucous cells cause a relocation of serous cells to the endpiece periphery. This concept has been uncritically accepted by several authors of histology textbooks. This note presents a compendium of observations and unanswered questions that render this proposition untenable.

Objective: The aim of our study was to establish a protocol to quantify the glycogen from human platelets. Methods: The total glycogen was determined in the platelet concentrate (PC) obtained from healthy male volunteers. The assay comprised the following steps: (1) cell lysing and glycogen extraction, (2) precipitation of glycogen and its washing, (3) conversion of glycogen to free glucose, (4) neutralization, and (5) detection of free glucose. Results: In our analysis, the measured glycogen content in the PC was 0.263 ± 0.101 μg/106 platelets (mean ± SD, n=16). Discussion: Despite the potential importance of glycogen determination in the platelets, a biochemical method to quantify this polymer in these cells is not available at the moment. Our protocol proved to be a simple method to measure these reserves from a blood sample. We expect to contribute to future studies of platelet metabolism, especially regarding glycogen function in these cells.

Molecular testing for EGFR mutations and ALK gene rearrangements has become a routine part of lung cancer pathological diagnosis, and is critical to determine the most effective therapies for patients with this poor prognosis cancer. ALK gene rearrangements are seen particularly in adenocarcinoma of the lung and are associated with an excellent response rate to targeted inhibition with crizotinib in clinical trials for many patients whose tumours harbour this change. ALK gene rearrangement is generally observed at low incidence posing challenges for routine detection. Since a high proportion of lung cancers are inoperable at presentation, cytology plays a central role in diagnosis and provision of material for ALK testing. The advantages offered by cytology specimens for ALK and other molecular testing are becoming increasingly recognised - cytology specimens tend to have a lower proportion of contaminating stromal and other non-neoplastic cells and often have higher quality DNA than routine histology specimens. One challenge is the often limited amount of cytological material obtaining in many lung cancer cytology specimens. FISH testing for ALK gene rearrangement using a break-apart probe is the gold standard for testing although there is a strong role for immunohistochemistry in ALK testing. This review highlights key aspects of ALK testing in cytology specimens.

The discovery of diabetes mellitus (simplified as “diabetes”) could be traced back to 1500 B.C. The first detailed description appeared in 1800, but in the last few decades, the prevalence of diabetes increases dramatically. Till present, according to WHO’s report, 247 million people worldwide have been diagonized Diabetes, among which 3.47 million died from diabetes. Thus, WHO projects that diabetes will become the 7th leading cause of death in 2030. The typical feature of diabetes is hyperglycemia as a result of metabolic disorder. As the disease progress, complications develop including macrovascular and microvascular abnormality, leading to multiple organ failure and mortality. Apart from diabetes-associated vascular diseases, numerous studies have documented the association between hyperglycemia and cancer. There are three main types of diabetes: (1) type 1 diabetes mellitus (T1DM) in which pancreatic β cell death by immune attack; (2) type 2 diabetes mellitus (T2DM) in which pancreatic β cells fail to produce sufficient amount of insulin; and (3) gestational diabetes which occurs in pregnant women due to high demand for insulin production. Type 2 DM accounts for 80-90% of diabetes and obesity has been identified as a strong and modifiable risk factor in type 2 DM. In this review, we first provide a general introduction of T2DM in the aspects of diagnosis, pathogenesis and clinical characteristics of T2DM. We will focus on how obesity has the adverse impact on insulin resistance, pancreatic β cell death and dysfunction and the development of T2DM. Next, we will summarize the animal models of T2DM including their advantages and drawbacks when compared to their clinical relevance. Finally, we will summarize the interventions that have been applied to treat obesity and T2DM and the remaining problems.

Objective: To elucidate the mechanism of formation of calcific deposits inside the human natural heart valve based on their ultra-fine structure on nanometer scale observed by atomic force microscope.
Methods: Cross-sections of an aortic valve calcification were observed by scanning electron microscope. Thin slices several micrometeres thick from the same deposit were observed by atomic force microscopy in the Peak Force Imagining mode providing topographic, adhesion and quantitative nanoscale elastic modulus maps of the surface.
Results: Most of the aortic valve calcification was composed of large blocks of mostly compact matter, porous on the microscopic scale. The mineral blocks consisted mainly of closely arranged elongated needle- and plate-like crystals, 30 to 70 nm in diameter, and irregularly disseminated areas of soft organic material.
Conclusions: Crystals forming mineral blocks of mostly compact matter are nucleated on organic substrates, with their growth controlled by the diffusion of building units through the virtually stagnant layer of interstitial fluid wetting the deposit surface. Preferentially formed precursors of hydroxyapatite, dicalcium, octacalcium and/or amorphous calcium phosphate, are in later stages of deposit development transformed into biological hydroxyapatite.

Background and objectives: Methotrexate (MTX) had been used for many years and complications usually encountered during treatment especially in cancer patients. The aim of the present study was to determine the preventive and early-stage anti cytotoxic effects of green tea on the histology of the submandibular gland of rats treated by high single dose of methotrexate.
Materials and methods: The study included 36 Albino rats. Twelve animals were used in the pilot study to find the maximum toxic dose, and the other twenty four were divided into four groups, vehicle treated control group, green tea extract treated control group (40 mg/kg/day), methotrexate treated group (80 mg/kg), and methotrexate and green tea extract treated group. Submandibular glands excisions were then performed. Histopathological examination was performed with hematoxylin-eosin, Masson’s trichrome, and PAS stains. Cell proliferation was examined using the Ki-67 antibody and anti apoptotic effect was determined based on Bcl-2 staining.
Results: In the methotrexate and green tea extract treated group a non significant difference in the Ki-67 expression (p>0.01) and a significant increase in Bcl-2 expression (p<0.01) were seen in comparison with the methotrexate treated group.
Conclusion: Green tea aqueous extract at a concentration of 40 mg/kg/day produced a protection against methotrexate induced cytotoxicity in rat submandibular gland by increasing the expression of Bcl-2.

Liver abscess was a bacterial infected disease. Liver abscess could be described as Gas-forming and non-Gasforming. There was high prevalence of liver abscess in Asia, especially in Taiwan, Singapore and Korea. The patient was a 48-year-old Taiwanese woman who admitted to our hospital with abdominal pain for 1 week. After abdominal computed tomography (CT) scan, a large gas forming abscess was presented (15.7 × 13.6 × 17.6 cm). Klebsiella pneumoniae was cultured from pus and blood. The patient was treated with antibiotics and status post (s/p)-pig tail drainage for two times. Gas-forming liver abscess was a disease with high mortality rate and induced bacteremia. Therefore, Gas-forming liver abscess was thought important and has to be treated immediately

The aim of this study was to show how optical coherence tomography (OCT) can provide means for assessing macula and the optic disc, and so can be used to diagnose and follow micro-structural, macular, changes (epiretinal membranes; macular edema; lamellar hole and discontinuity of inner/outer segment line), that occur as complications of a disease (i.e. uveitis). It is a non invasive, non contact method that provides cross-sectional images of retina and its substructures in a real-time mode. High definition cross-sectional images obtained by a commercially available OCT are attained at 1-15 μm and can provide detailed pictures from deep within the retina. Seeing that OCT can be repeated numerous times it is useful for practitioners that need to follow treatment effects and progression of the disease. Spectral-Domain OCT was used to diagnose and follow macular changes in patients with uveitis (intermediate and posterior) and retinal vasculitis. OCT showed an epiretinal membrane (ERM) as intense back-reflective layer on the inner retinal surface. Via this method thickness measurements of ERM as well as the degree of inner retinal surface attachment (focal or diffused) were acquired and precisely defined. OCT showed macular edema as retinal layering followed by presence of intraretinal cavities and absence of back-reflection. It provided means of measuring thickness, localization, as well as size of intraretinal cysts during the evolution of macular edema (ME), and so was utilized in analysis of various factors such as progression of ERM; presence of ME; surface roughness; changes of thickness and layer configuration of retinal tissue. OCT macular scan provides precise and quick diagnosis of delicate microstructural retinal changes in high definition that can’t be obtained during classical clinical examination. Progression of disease, changes and treatment effects, followed via OCT, are of great importance in clinical practice.

The aim of this study is to investigate the effect of four herbal extract and their efficacy on the histomorphometry of the kidney in STZ induced diabetic rats with a view to understanding their antidiabetic properties. Forty- two healthy adult Wistar rats (Rattus novergicus) with an average weight of 153.4 g were randomly divided into seven groups (n=6). STZ (65 mg/kg) dissolved in citrate buffer was administered intraperitoneally to animals in groups (B-G) while animals in group A received equivalent volume of citrate buffer. Plant extracts (100 mg/kg) were administered daily (orally) to animals in groups C-F and glimepiride (anti-diabetic drug) to animals in group G for fourteen days. After the expiration of the study the animals were sacrificed and the kidneys were excised, fixed in 10% formol saline for histology and morphometric analysis. The glomeruli of the diabetic group were atrophied which is validated by significant decrease in its density, shrinkage and increased bowman’s space. These observations were also characterized by diminished cellular proliferation, decreased cellular volume and ischemia. The histology and morphometric analysis revealed that the kidney in the group treated with Psidium guajava shows a better histoarchitectural outline of all the four plant extracts used. This present study therefore suggests that Psidium guajava could be a better alternative therapy in ameliorating diabetic-associated disorders of the kidney.