Adult ; Cerebral Ventricle Neoplasms ; diagnosis ; metabolism ; Diagnosis, Differential ; Glial Fibrillary Acidic Protein ; analysis ; Humans ; Immunohistochemistry ; Lateral Ventricles ; chemistry ; pathology ; Male

OBJECTIVETo evaluate the efficiency of a modified culture method for rat cerebral cortical astrocytes in vitro.

METHODSThe astrocytes derived from the cerebral cortex of 3-day-old Sprague-Dawley rats were first purified as described previously, then the cells were replanted at a low density. The culture flask was changed after 1 hour and substratum was replaced after 24 hours. Cells were syncretized to a monolayer, followed by cell passage. After three passages the cells were cultured in DMEM medium containing 10% fetal serum for a long period. The derivation of the cells was identified by immunofluorescent staining with anti-GFAP polyclonal antibodies.

RESULTSA variety of morphologically distinct astrocytes with many long processes and small cell bodies were obtained. Finally an astrocytic network occurred through cellular process connections. The immunofluorescent staining demonstrated the percentage of GFAP-positive cells was above 98%.

CONCLUSIONSThe modified culture method for astrocytes from rat cerebral tissue is reliable, with a high purity. The cultured astrocytes have a similar morphological development to those in vivo.

Animals ; Astrocytes ; physiology ; Cell Culture Techniques ; Cerebral Cortex ; cytology ; Female ; Glial Fibrillary Acidic Protein ; analysis ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study influencing factors of detection of bovine central nervous system (CNS) tissue contaminated beef by enzyme immunoassay (EIA), and the method was applied to the detection of imported beef and domestic beef of China.

METHODSRaw beef homogenates containing different concentrations of raw CNS tissue and the same samples which were heated were detected after different time by RIDASCREEN(r) Risk Material 10/5 and RIDASCREEN(r) Probennahme- zubehor Sampling tools kits. PBS suspension and sample dilution buffer (SDB) suspension of bovine brain tissue with the same concentration of the standard were detected. Beef from USA and domestic market of China were then detected by the kits.

RESULTSThe kits could detect both raw and heated CNS tissue in the products with high sensitivity. The absorbance values (AV) increased with the concentrations of CNS in samples. Heating and increasing of time could decrease the absorbance values of the samples which contain CNS tissue. The AV of the PBS suspension of bovine brain tissue was higher than the SDB suspension and the AV of both were higher than the AV of standard of the same concentration. No CNS tissue was detected from all imported beef. No CNS tissue was detected in all samples from domestic market of China except for foxtail.

CONCLUSIONThe EIA method has high sensitivity for detection of bovine CNS tissue contaminated beef with the glial fibrillary acidic protein (GFAP) as accurate target substance. Heating and increasing of time can lead to decreasing of the AV of samples. Improper slaughter process can lead to contamination of bovine products by bovine CNS tissue.

Animals ; Brain ; metabolism ; Brain Chemistry ; Cattle ; Food Contamination ; analysis ; Food Inspection ; methods ; Glial Fibrillary Acidic Protein ; analysis ; Immunoenzyme Techniques ; Meat Products ; analysis

OBJECTIVETo investigate the molecular basis of infantile Alexander disease in a Chinese patient, which may yield useful information for further genetic counseling.

METHODSDNA sequencing analysis and restriction endonuclease analysis were used to detect the mutation of glial fibrillary acidic protein (GFAP) gene in a patient with clinically diagnosed Alexander disease, in her parents and in 50 healthy controls.

RESULTSA 249C>T (R79C) mutation was identified in the exon 1 of the GFAP gene but not in her parents and the controls.

CONCLUSIONThe study on mutation of GFAP gene in Chinese patients with Alexander disease has never been reported previously. The mutation analysis of GFAP gene can provide valuable information for the diagnosis of Alexander disease and can serve as a reliable method of prenatal diagnosis for the family.

Alexander Disease ; diagnosis ; genetics ; Base Sequence ; Child, Preschool ; China ; DNA Mutational Analysis ; Female ; Genetic Predisposition to Disease ; Glial Fibrillary Acidic Protein ; genetics ; Humans ; Mutation ; Polymerase Chain Reaction

Recently the origin of gastrointestinal stromal tumors (GISTs) is thought be the interstitial cells of Cajal or primitive stem cells. This study was performed to evaluate the roles of fine needle aspiration cytology (FNAC), cell block preparation, and immunohistochemistry in the diagnosis of GISTs. Nine cases of GIST in which FNAC was performed were included in this study. Cytologically, the tumor cells characteristically occurred in closely packed cohesive tissue fragments with high cellular density often in bloody background. The tumor cells often formed fascicles with parallel, side-by-side arrangements of the nuclei. Histologically, GISTs were highly cellular spindle or epithelioid tumor with basophilic appearance. Immunohistochemically, GISTs were c-kit positive in all of nine cases, CD34 positive in seven, focally SMA positive in two, and S-100 and GFAP negative in all. Both histologic and cell block sections showed the same histologic and immunohistochemical features. Cytomorphologically GISTs show a broad morphologic spectrum but rarely a significant nuclear pleomorphism and the assessment of malignant potential is difficult based on cytology alone. However, in the appropriate clinical and radiologic setting, a confident diagnosis of primary or metastatic GIST can be established by FNAC, cell block, and immunohistochemistry.
Actins/analysis ; Adult ; Antigens, CD34/analysis ; Biopsy, Needle ; Female ; Gastrointestinal Neoplasms/chemistry/*pathology ; Glial Fibrillary Acidic Protein/analysis ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Paraffin Embedding ; Proto-Oncogene Proteins c-kit/analysis ; S100 Proteins/analysis ; Stromal Cells/*pathology

OBJECTIVEThe purpose of this study was to culture and identify neural stem cells from mouse embryos in vitro using a modified method and provide a basis for further study of the biology of neural stem cells under hypoxia.

METHODSThe cells were isolated mechanically from the front cortex of fetal Institute of Cancer Research (ICR) mice on embryonic day 14. They were passaged by mechanical dissociation and enzymatic digestion. The neurospheres were identified by immunofluorescent staining of nestin. Cell differentiation was induced by 1% fetal bovine serum and then the cells were identified by immunohistochemistry of β-tubulin III and GFAP.

RESULTSThe cells obtained from the front cortex of fetal ICR mice had the capacity of forming neurospheres which showed nestin immunoreactive positivity. After being induced by 1% fetal bovine serum, the cells were differentiated into β-tubulin III-positive cells and GFAP-positive cells.

CONCLUSIONSUsing mechanical dissociation of primary cells and mechanical dissociation with enzymatic digestion of primary cells, the NSCs from the front cortex of mouse embryos can be obtained.

Animals ; Cell Differentiation ; Cells, Cultured ; Embryo, Mammalian ; cytology ; Female ; Glial Fibrillary Acidic Protein ; analysis ; Intermediate Filament Proteins ; analysis ; Mice ; Mice, Inbred ICR ; Nerve Tissue Proteins ; analysis ; Nestin ; Neural Stem Cells ; chemistry ; cytology ; Tubulin ; analysis

BACKGROUNDFunctional electrical stimulation (FES) is known to promote the recovery of motor function in rats with ischemia and to upregulate the expression of growth factors which support brain neurogenesis. In this study, we investigated whether postischemic FES could improve functional outcomes and modulate neurogenesis in the subventricular zone (SVZ) after focal cerebral ischemia.

METHODSAdult male Sprague-Dawley rats with permanent middle cerebral artery occlusion (MCAO) were randomly assigned to the control group, the placebo stimulation group, and the FES group. The rats in each group were further assigned to one of four therapeutic periods (1, 3, 7, or 14 days). FES was delivered 48 hours after the MCAO procedure and divided into two 10-minute sessions on each day of treatment with a 10-minute rest between them. Two intraperitoneal injections of bromodeoxyuridine (BrdU) were given 4 hours apart every day beginning 48 hours after the MCAO. Neurogenesis was evaluated by immunofuorescence staining. Wnt-3 which is strongly implicated in the proliferation and differentiation of neural stem cells (NSCs) was investigated by Western blotting analysis. The data were subjected to one- way analysis of variance (ANOVA), followed by a Tukey/Kramer or Dunnett post hoc test.

RESULTSFES significantly increased the number of BrdU-positive cells and BrdU/glial fibrillary acidic protein double- positive neural progenitor cells in the SVZ on days 7 and 14 of the treatment (P < 0.05). The number of BrdU/doublecortin (DCX) double-positive migrating neuroblast cells in the ipsilateral SVZ on day 14 of the FES treatment group ((522.77 ± 33.32) cells/mm(2)) was significantly increased compared with the control group ((262.58 ± 35.11) cells/mm(2), P < 0.05) and the placebo group ((266.17 ± 47.98) cells/mm(2), P < 0.05). However, only a few BrdU/neuron-specific nuclear protein-positive cells were observed by day 14 of the treatment. At day 7, Wnt-3 was upregulated in the ipsilateral SVZs of the rats receiving FES ((0.44 ± 0.05)%) compared with those of the control group rats ((0.31 ± 0.02)%, P < 0.05) or the placebo group rats ((0.31 ± 0.04)%, P < 0.05). At day 14, the corresponding values were (0.56 ± 0.05)% in the FES group compared with those of the control group rats ((0.50 ± 0.06)%, P < 0.05) or the placebo group rats ((0.48 ± 0.06)%, P < 0.05).

CONCLUSIONFES augments the proliferation, differentiation, and migration of NSCs and thus promotes neurogenesis, which may be related to the improvement of neurological outcomes.

Animals ; Bromodeoxyuridine ; metabolism ; Cell Proliferation ; Cerebral Ventricles ; physiopathology ; Electric Stimulation Therapy ; Glial Fibrillary Acidic Protein ; analysis ; Male ; Neural Stem Cells ; physiology ; Neurogenesis ; Rats ; Rats, Sprague-Dawley ; Stroke ; physiopathology ; therapy ; Wnt3A Protein ; analysis

OBJECTIVETo study the effect of Danshensu (DSS) on directional differentiation of mesenchymal stem cells (MSC) into neuron-like cells.

METHODSMSC were separated from bone marrow with density gradient centrifugation, wall sticking screening and amplified in vitro. Flow cytometry was used to monitor the expression of surface antigens. DSS contained in non-serum L-DMEM was used to induce differentiation of MSC to neuronlike cells, and the effect of DSS when different concentration and acting time used was explored. And levels of neuron-specific enolase (NSE), neurofilament protein (NF-M), nestin, and expression of glial fibrillary acidic protein (GFAP) were measured by immunohistochemical method.

RESULTSAfter being propagated and amplified in vitro, MSC were positively expressed for CD29, CD44, CD166, and negatively expressed for CD14, CD34, CD45, HLA-DR. After induction of DSS, MSC exhibited the typical form of perikaryon with pyknotic cell body and prominence projected like that of neuron. These cells were positively expressed in NSE, NF-M and nestin, and negatively expressed in GFAP.

CONCLUSIONDSS could induce differentiation of MSC to neuron-like cells in vitro, the action is concentration- and time-dependent.

Bone Marrow Cells ; cytology ; Cell Differentiation ; Cell Division ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Glial Fibrillary Acidic Protein ; analysis ; Humans ; Lactates ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; Neurofilament Proteins ; analysis ; Neurons ; cytology ; Phosphopyruvate Hydratase ; analysis

BACKGROUNDThe two most basic properties of mesenchymal stem cells (MSCs) are the capacities to self-renew indefinitely and differentiate into multiple cells and tissue types. The cells from human umbilical cord Wharton's Jelly have properties of MSCs and represent a rich source of primitive cells. This study was conducted to explore the possibility of inducing human umbilical cord Wharton's Jelly-derived MSCs to differentiate into nerve-like cells.

METHODSMSCs were cultured from the Wharton's Jelly taken from human umbilical cord of babies delivered after full-term normal labor. Salvia miltiorrhiza and beta-mercaptoethanol were used to induce the human umbilical cord-derived MSCs to differentiate. The expression of neural protein markers was shown by immunocytochemistry. The induction process was monitored by phase contrast microscopy, electron microscopy (EM), and laser scanning confocal microscopy (LSCM). The pleiotrophin and nestin genes were measured by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSMSCs in the Wharton's Jelly were easily attainable and could be maintained and expanded in culture. They were positive for markers of MSCs, but negative for markers of hematopoietic cells and graft-versus-host disease (GVHD)-related cells. Treatment with Salvia miltiorrhiza caused Wharton's Jelly cells to undergo profound morphological changes. The induced MSCs developed rounded cell bodies with multiple neurite-like extensions. Eventually they developed processes that formed networks reminiscent of primary cultures of neurons. Salvia miltiorrhiza and beta-mercaptoethanol also induced MSCs to express nestin, beta-tubulinIII, neurofilament (NF) and glial fibrillary acidic protein (GFAP). It was confirmed by RT-PCR that MSCs could express pleiotrophin both before and after induction by Salvia miltiorrhiza. The expression was markedly enhanced after induction and the nestin gene was also expressed.

CONCLUSIONSMSCs could be isolated from human umbilical cord Wharton's Jelly. They were capable of differentiating into nerve-like cells using Salvia miltiorrhiza or beta-mercaptoethanol. The induced MSCs not only underwent morphologic changes, but also expressed the neuron-related genes and neuronal cell markers. They may represent an alternative source of stem cells for central nervous system cell transplantation.

Cell Differentiation ; Cells, Cultured ; Glial Fibrillary Acidic Protein ; analysis ; Humans ; Immunohistochemistry ; Mesenchymal Stromal Cells ; cytology ; Neurofilament Proteins ; analysis ; Neurons ; cytology ; Reverse Transcriptase Polymerase Chain Reaction ; Tubulin ; analysis ; Umbilical Cord ; cytology

We report a case of carcinoma ex pleomorphic adenoma of a sublingual gland in a 70-year-old man. Under a clinical diagnosis of benign salivary gland tumor, excision of the mass with the sublingual salivary gland in an en bloc fashion via an intraoral approach was performed. Histopathologically, there was a rupture of the fibrous capsule and diffuse cell-rich sheets composed of myoepithelial cells with round nuclei were also seen. Immunohistochemically, the cells that composed of cell rich sheets were positive to smooth muscle actin. Final diagnosis of myoepithelial carcinoma ex pleomorphic adenoma was made.
Adenoma, Pleomorphic ; pathology ; Aged ; Carcinoma ; chemistry ; pathology ; Diagnosis, Differential ; Glial Fibrillary Acidic Protein ; analysis ; Humans ; Keratins ; analysis ; Male ; Myoepithelioma ; chemistry ; pathology ; Neoplasm Invasiveness ; S100 Proteins ; analysis ; Sublingual Gland Neoplasms ; chemistry ; pathology