OBJECTIVE: To estimate the detrimental effects of shortwave exposure on rat hippocampal structure and function and explore the underlying mechanisms. METHODS: One hundred Wistar rats were randomly divided into four groups (25 rats per group) and exposed to 27 MHz continuous shortwave at a power density of 5, 10, or 30 mW/cm2 for 6 min once only or underwent sham exposure for the control. The spatial learning and memory, electroencephalogram (EEG), hippocampal structure and Nissl bodies were analysed. Furthermore, the expressions of N-methyl-D-aspartate receptor (NMDAR) subunits (NR1, NR2A, and NR2B), cAMP responsive element-binding protein (CREB) and phosphorylated CREB (p-CREB) in hippocampal tissue were analysed on 1, 7, and 14 days after exposure. RESULTS: The rats in the 10 and 30 mW/cm2 groups had poor learning and memory, disrupted EEG oscillations, and injured hippocampal structures, including hippocampal neurons degeneration, mitochondria cavitation and blood capillaries swelling. The Nissl body content was also reduced in the exposure groups. Moreover, the hippocampal tissue in the 30 mW/cm2 group had increased expressions of NR2A and NR2B and decreased levels of CREB and p-CREB. CONCLUSION Shortwave exposure (27 MHz, with an average power density of 10 and 30 mW/cm2) impaired rats' spatial learning and memory and caused a series of dose-dependent pathophysiological changes. Moreover, NMDAR-related CREB pathway suppression might be involved in shortwave-induced structural and functional impairments in the rat hippocampus.
Animals ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Dose-Response Relationship, Radiation ; Electroencephalography ; radiation effects ; Hippocampus ; radiation effects ; Male ; Memory ; radiation effects ; Nissl Bodies ; physiology ; radiation effects ; Radio Waves ; adverse effects ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, N-Methyl-D-Aspartate ; genetics ; metabolism ; Spatial Learning ; radiation effects

Cerebrospinal fluid (CSF) contains several molecules which are essential for neurogenesis. Human dental pulp stem cells (hDPSCs) are putatively neural crest cell-derived that can differentiate into neurons and glial cells under appropriate neurotrophic factors. The aim of this study was to induce differentiation of hDPSCs into neuroglial phenotypes using retinoic acid (RA) and CSF. The hDPSCs from an impacted third molar were isolated by mechanical and digestion and cultured. The cells have treated by 10⁻⁷µM RA (RA group) for 8 days, 10% CSF (CSF group) for 8 days and RA with CSF for 8 days (RA/CSF group). Nestin, microtubule-associated protein 2 (MAP2), and glial fibrillary acidic protein immunostaining were used to examine the differentiated cells. Axonal outgrowth was detected using Bielschowsky's silver impregnation method and Nissl bodies were stained in differentiated cells by Cresyl violet. The morphology of differentiated cells in treated groups was significantly changed after 3–5 days. The results of immunocytochemistry showed the presence of neuroprogenitor marker nestin was seen in all groups. However, the high percentage of nestin positive cells and MAP2, as mature neural markers, were observed at the pre-induction and induction stage, respectively. Nissl bodies were detected as dark-blue particles in the cytoplasm of treated cells. Our findings showed the RA as pre-inducer and CSF as inducer for using in vitro differentiation of neuron-like cells and neuroglial cells from hDPSCs.
Axons ; Cerebrospinal Fluid* ; Cytoplasm ; Dental Pulp* ; Digestion ; Glial Fibrillary Acidic Protein ; Humans* ; Immunohistochemistry ; In Vitro Techniques ; Methods ; Microtubule-Associated Proteins ; Molar, Third ; Nerve Growth Factors ; Nestin ; Neural Crest ; Neurogenesis ; Neuroglia* ; Neurons ; Nissl Bodies ; Phenotype ; Silver ; Stem Cells* ; Tretinoin* ; Viola

This study was carried out to explore the effect of DNA hypomethylation on chondrocytes phenotype, in particular the effect on chondrocyte hypertrophy, maturation, and apoptosis. Chondrocytes derived from caudal region of day 17 embryonic chick sterna were pretreated with hypomethylating drug 5-aza-2'-deoxycytidine for 48 hours and then maintained in the normal culture medium for up to 14 days. Histological studies showed distinct morphological changes occurred in the pretreated cultures when compared to the control cultures. The pretreated chondrocytes after 7 days in culture became bigger in size and acquired more flattened fibroblastic phenotype as well as a loss of cartilage specific extracellular matrix. Scanning electron microscopy at day 7 showed chondrocytes to have increased in cell volume and at day 14 in culture the extracellular matrix of the pretreated cultures showed regular fibrillar structure heavily embedded with matrix vesicles, which is the characteristic feature of chondrocyte hypertrophy. Transmission electron microscopic studies indicated the terminal fate of the hypertrophic cells in culture. The pretreated chondrocytes grown for 14 days in culture showed two types of cells: dark cells which had condense chromatin in dark patches and dark cytoplasm. The other light chondrocytes appeared to be heavily loaded with endoplasmic reticulum indicative of very active protein and secretory activity; their cytoplasm had large vacuoles and disintegrating cytoplasm. The biosynthetic profile showed that the pretreated cultures were actively synthesizing and secreting type X collagen and alkaline phosphatase as a major biosynthetic product.
Alkaline Phosphatase ; Apoptosis ; Cartilage ; Cell Size ; Chondrocytes* ; Chromatin ; Collagen Type X ; Cytoplasm ; DNA ; Endoplasmic Reticulum ; Endoplasmic Reticulum, Rough ; Extracellular Matrix ; Fibroblasts ; Hypertrophy* ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Phenotype ; Vacuoles

The success of regeneration attempt is based on an ideal combination of stem cells, scaffolding and growth factors. Tissue constructs help to maintain stem cells in a required area for a desired time. There is a need for easily obtainable cells, potentially autologous stem cells and a biologically acceptable scaffold for use in humans in different difficult situations. This study aims to address these issues utilizing a unique combination of stem cells from gingiva and a hydrogel scaffold, based on a natural product for regenerative application. Human gingival mesenchymal stem cells (HGMSCs) were, with due induction, differentiated to neuronal lineages to overcome the problems associated with birth tissue-related stem cells. The differentiation potential of neuronal lineages was confirmed with suitable specific markers. The properties of mesenchymal stem cells in encapsulated form were observed to be similar to free cells. The encapsulated cells (3D) were then subjected to differentiation into neuronal lineages with suitable inducers, and the morphology and gene expression of transient cells were analyzed. HGMSCs was differentiated into neuronal lineages as both free and encapsulated forms without any significant differences. The presence of Nissl bodies and the neurite outgrowth confirm the differentiation. The advantages of this new combination appear to make it a promising tissue construct for translational application.
Gene Expression ; Gingiva ; Humans* ; Hydrogel* ; Intercellular Signaling Peptides and Proteins ; Mesenchymal Stromal Cells* ; Neurites ; Neurons* ; Nissl Bodies ; Parturition ; Regeneration ; Stem Cells

PURPOSE: Electric injury may result in cataracts, which usually occur bilaterally. In this report, we present a rare complication of such an injury presenting as a unilateral cataract with optic neuropathy. CASE SUMMARY: A 17-year-old male patient presented with gradual worsening of vision in his right eye 7 weeks after sustaining an injury from 22,900 volts of high-voltage electric current his right hand. On initial eye examination, the best corrected visual acuity (BCVA) was 20/60 in the right eye and 20/20 in the left eye. Relative afferent pupillary defect (RAPD) was noted in the right eye. Slit-lamp examination confirmed the typical anterior subcapsular lenticular opacities and funduscopy showed a slightly pale optic nerve head in the right eye. Pattern reversal visually evoked potential (P-VEP) showed a significant delay of P-100 implicit time in the right eye. After 7 months, phacoaspiration and posterior chamber intraocular lens implantation were performed in the right eye. Postoperatively, the BCVA improved to 20/30 but visual field examination showed a partial defect in the temporal area of the right eye. Cataract did not develop in the left eye during 15 months of follow-up. The electron microscopic findings showed that the number of mitochondria in the cytoplasm increased. The rough endoplasmic reticulum in the cytoplasm and microfilaments were enriched. CONCLUSIONS: Manifestation time of ocular complications after electric injuries is different. Therefore an ophthalmic examination should be performed regularly in the early recovery period of such injuries and in particular, progression of bilateral electric cataract must be checked.
Actin Cytoskeleton ; Cataract ; Cytoplasm ; Electric Injuries ; Electrons ; Endoplasmic Reticulum, Rough ; Evoked Potentials ; Eye ; Follow-Up Studies ; Hand ; Humans ; Lens Implantation, Intraocular ; Male ; Mitochondria ; Optic Disk ; Optic Nerve Diseases ; Pupil Disorders ; Vision, Ocular ; Visual Acuity ; Visual Fields

OBJECTIVETo study the effect of erythropoietin (EPO) on the activities of antioxidant enzymes, namely catalase (CAT) and glutathione peroxidase (GSH-Px) in the brain tissues of aged rats.

METHODSThirty SD rats were randomly divided into normal control, aging model, and recombinant human erythropoietin (rhEPO) treatment groups (n=10). Morris water maze was used to compare the behavioral indexes. The rats were then sacrificed to observe Nissl bodies in the hippocampal neurons with Nissl staining and test the activities of CAT and GSH-Px in the brain tissues.

RESULTSCompared with the control group, the aging rats showed significantly deteriorated learning and memory abilities (P<0.05), which were improved obviously by rhEPO treatment (P<0.05). The number of Nissl bodies in the neurons was reduced in the aging rats compared with that in the control group, and rhEPO treatment increased the number of Nissle bodies but failed to restore the control level. The aging rats also showed significantly lowered activities of CAT and GSH-Px in the brain tissue (P<0.05), which were increased significantly after rhEPO treatment (P<0.05).

CONCLUSIONEPO can enhance the activities of the antioxidant enzymes in the brain tissues of aged rats to increase the antioxidant capacity and produces an anti-aging effect.

Aging ; Animals ; Brain ; enzymology ; Catalase ; metabolism ; Epoetin Alfa ; Erythropoietin ; pharmacology ; Glutathione Peroxidase ; metabolism ; Learning ; drug effects ; Male ; Memory ; drug effects ; Nissl Bodies ; drug effects ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; pharmacology

Transient bullous dermolysis of the newborn (TBDN) is a rare subtype of the dystrophic epidermolysis bullosa characterized by blistering at birth which improves spontaneously during early life. Electron microscopy showed sublamina densa separation with dilated rough endoplasmic reticulum and electron dense inclusions. Immunofluorescence mapping using anti-type VII collagen antibody showed widespread intraepidermal type VII collagens which are a characteristic finding of TBDN. Here, we report two cases of TBDN presenting typical clinical manifestations, electron microscopy findings, and immunofluorescence mapping results. The skin lesions of both patients healed spontaneously 2~3 months later.
Blister ; Collagen ; Collagen Type VII ; Electrons ; Endoplasmic Reticulum, Rough ; Epidermolysis Bullosa Dystrophica ; Fluorescent Antibody Technique ; Humans ; Infant, Newborn ; Microscopy, Electron ; Parturition ; Skin

A 37-year-old female presented with a conjunctival mass discovered 3 years prior. An excisional biopsy revealed a patternless proliferation of round and spindle-shaped cells with an eosinophilic fibrillary cytoplasm and vesicular nuclei with occasional inclusions. Psammoma bodies were arranged around the dilated irregularly-shaped vessels. Differential diagnoses included conjunctival solitary fibrous tumor (SFT), nevus, glomangioma, ectopic meningioma, and mesectodermal leiomyoma. The tumor cells were immunoreactive for CD34, CD99, bcl-2 and vimentin, and were negative for smooth muscle actin, desmin, glial fibrillary acidic protein, S-100 protein, epithelial membrane antigen, and human melanoma black-45. Ultrastructurally, the tumor cells had rough endoplasmic reticulum, free ribosomes, and scattered mitochondria without basal lamina or cellular junctions, which are features of fibroblasts. A diagnosis of SFT was rendered based on the light microscopic, immunohistochemical, and electron microscopic findings. We report here on the second case of a SFT arising in the conjunctiva, which clinically and histologically mimics conjunctival nevus, glomangioma, ectopic meningioma, and a hybrid neurogenic-myogenic tumor such as mesectodermal leiomyoma.
Actins ; Adult ; Antigens, CD34 ; Basement Membrane ; Biopsy ; Chimera ; Conjunctiva ; Cytoplasm ; Desmin ; Diagnosis, Differential ; Electrons ; Endoplasmic Reticulum, Rough ; Eosinophils ; Female ; Fibroblasts ; Glial Fibrillary Acidic Protein ; Glomus Tumor ; Humans ; Leiomyoma ; Light ; Melanoma ; Meningioma ; Mitochondria ; Mucin-1 ; Muscle, Smooth ; Nevus ; Ribosomes ; S100 Proteins ; Solitary Fibrous Tumors ; Vimentin

AIMTo study the effects of noise on event-related potential(ERP) and its mechanism in hippocampus in rats.

METHODSMale SD rats were divided into 2 groups: control group (CG) and noise exposure group(NG). The rats in NG were exposed to white noise 105 dB SPL for 2.5 h/d x 20 d. P300 were recorded at parietal bone in rats. The Nissl body, NMDAR2B and [Ca2+]i of neurons in hippocampus were analyzed.

RESULTSThe peak latency (PL) of ERP P3a, P3 and P3b in NG were significantly longer than that in CG in the 14th and 20th exposure day. The amount of Nissl body in dentate gyrus (DG) and CA1 region and NMDAR2B in DG, CA1 and CA3 region of hippocampus of NG were significantly decreased than those of CG as well, while the concentration of Ca2+ in neurons increased markedly in NG.

CONCLUSIONDecreased Nissl body and NMDAR2B and increased [Ca2+]i in hippocampus in long-term noise exposed rats might cause the change of ERP P300.

Animals ; Calcium ; metabolism ; Environmental Exposure ; adverse effects ; Event-Related Potentials, P300 ; physiology ; Hippocampus ; metabolism ; physiology ; Male ; Neurons ; metabolism ; physiology ; Nissl Bodies ; metabolism ; Noise ; adverse effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism

OBJECTIVETo observe the ultrastructural changes of HeLa cells in response to tubeimoside I (TBMS1) treatment and the protective effect of cyclosporine A (CsA), and explore the role of intrinsic apoptosis pathway in TBMS1-induced HeLa cell apoptosis.

METHODSHeLa cells were treated with TBMS1 (10-50 micromo/L) alone or in combination with 2 micromol/L CsA for 12 and 24 h and observed with transmission electron microscope (TEM) for the ultrastructural changes of the cells.

RESULTSTBMS1 induced apoptosis of HeLa cells in a concentration- and time-dependant manner. Under TEM, the treated cells progressively shrunk and the intercellular space widened with loss of microvillus, mitochondrial swelling, rough endoplasmic reticulum enlargement, chromatin condensation, nuclear shrinkage and nuclear pyknosis as TBMS1 concentration increased. At low concentrations, CsA offered partial protection of the mitochondria from TBMS1-induced damage whereas high-concentration CsA did not.

CONCLUSIONTBMS1 induces ultrastructural changes typical for apoptosis of the HeLa cells, which provides morphological evidence for the role of intrinsic apoptosis pathway in TBMS1-induced apoptosis.

Apoptosis ; drug effects ; Cell Nucleus ; drug effects ; ultrastructure ; Cyclosporine ; pharmacology ; Dose-Response Relationship, Drug ; Endoplasmic Reticulum, Rough ; drug effects ; ultrastructure ; Female ; HeLa Cells ; Humans ; Immunosuppressive Agents ; pharmacology ; Microscopy, Electron, Transmission ; Mitochondrial Swelling ; drug effects ; Saponins ; pharmacology ; Time Factors ; Triterpenes ; pharmacology ; Uterine Cervical Neoplasms ; pathology ; ultrastructure