OBJECTIVETo investigate the role of microtubule-associated protein 2 (MAP-2) and nestin in gastric development in human embryos and fetuses.

METHODSImmunohistochemistry was used to detect the expressions of MAP-2 and nestin proteins in the gastric cardia, pyloric and gastric tissues of human embryos and fetuses during the second, third and fourth month of development.

RESULTSIn the second to fourth months of gestation, MAP-2 and nestin expressions were detected in the neural cells and neural fibers of the intermuscular nerve plexus and submucosal plexus in the gastric cardia, pyloric and gastric tissues. As the gestational age increased, the number of MAP-2- and nestin-positive cells and the expression intensity all increased in the myenteric plexus, but MAP-2 and nestin expressions were negative in the glandular and mucosal tissues of human embryonic and fetal gastric cardia, pylorus or gastric walls.

CONCLUSIONMAP-2 and nestin participate in the regulation of the development of gastric tissues in human embryos.

Fetus ; metabolism ; Humans ; Intermediate Filament Proteins ; metabolism ; Microtubule-Associated Proteins ; metabolism ; Nerve Tissue Proteins ; metabolism ; Nestin ; Stomach ; embryology ; metabolism

OBJECTIVETo introduce the eukaryotic expression vector pEGFP-C1-PDX-1 into nestin-positive cell derived from bone marrow stromal cells by nucleofection and optimize the conditions for transfection.

METHODSThe recombinant plasmid was transfected into bone marrow stromal cells-derived nestin-positive cells with varied DNA quantities or the serum concentration in the medium. The expression of PDX-1 gene in the transfected cells was detected by RT-PCR.

RESULTSSatisfactory efficiency of transfection was achieved with the DNA quantity of 2-10 microg and medium serum concentration of 20%. PDX-1 expression was detected in the transfected cells by RT-PCR.

CONCLUSIONThe optimized transfection conditions result in enhanced efficiency of PDX-1 gene transfection into nestin-positive cells derived from bone marrow stromal cells, which may serve as the seed cells in tissue-engineering.

Bone Marrow Cells ; metabolism ; Genetic Vectors ; Homeodomain Proteins ; genetics ; Humans ; Intermediate Filament Proteins ; metabolism ; Nerve Tissue Proteins ; metabolism ; Nestin ; Plasmids ; Stromal Cells ; metabolism ; Trans-Activators ; genetics ; Transfection ; methods

OBJECTIVE: To observe the alteration of nestin intervals in the experimental traumatic brain injury and investigate its relation to the injury intervals. METHODS: The rat brain contusion was conducted by falling impact injury. After various survival interval (0.5, 6, 12 h and 1, 3, 7, 14, 28 d), immunohistochemical SP method was used for observing the expression of nestin in the cortex, hippocampal dentate gyrus and the corpus callosum on injury side. RESULTS: Expression of nestin positive cells increased at 0.5 h and reached the maximum level in 7 d after brain contusion, then the expression decreased gradually. The intensity of nestin staining in the the cortex and the hippocampal dentate gyrus decreased to normal on 28 d. As to the corpus callosum of injury side it remained weak on 28 d. CONCLUSION The changes of nestin immunohistochemical staining can be used as an index for forensic estimation of early injury time.
Animals ; Brain/metabolism* ; Brain Injuries/pathology* ; Cerebral Cortex/metabolism* ; Immunohistochemistry ; Intermediate Filament Proteins/metabolism* ; Male ; Nerve Tissue Proteins/metabolism* ; Nestin ; Rats ; Rats, Sprague-Dawley ; Staining and Labeling ; Time Factors

OBJECTIVETo investigate the effect of electroacupuncturing (EA) Hegu point (LI 4) on proliferation of neural stem cells in the subventricular zone (SVZ) after rat focal ischemia/reperfusion (I/R).

METHODSAnimal model was made by ligating external carotid artery and inserting a piece of nylon thread into the internal carotid artery. Eighty-eight Wistar rats were randomly divided into the normal control group (n = 4) ,the sham operated group (n =4),the IR groups (contain 1,7,14,21,28 days' five time points,each point n =8),and the IR + EA group (contain 1,7,14,21,28 days' five time points,each point n = 8). Bilateral Hegu points on rat were consecutively stimulated for 15 min,once daily, and 7 days as one treatment course. The protein expression of Nestin in SVZ was detected by immunochemistry method.

RESULTSCompared with the normal control group, the number of Nestin positive cell in SVZ in the I/R group increased strikingly on the 1st day, peaked on the 7 h day after reperfusion (P <0. 05), began to decrease on the 14th day, and almost reached normal on the 28 h day. A number of Nestin positive cells were present around the ischemic core in the cortex on the 7th day after reperfusion. After performed electroacupuncture, the number of Nestin positive cells in SVZ in the EA group increased on the 1st day, peaked on the 7th day, and increased 1. 5 times as compared with the IR 7 days group, the difference was obvious (P <0. 05) ,and began to decrease on the 14 h day, and almost reached normal on the 28th day. Nestin immunoreactivity around the ischemic core in the cortex was obviously presented in the EA 7 days group.

CONCLUSIONThe results indicate that cerebral ischemia can stimulate the potent of self-proliferation of neural stem cells and electroacupuncture can obviously increase the proliferation of neural stem cell.

Animals ; Cell Proliferation ; Electroacupuncture ; Intermediate Filament Proteins ; metabolism ; Medicine, Chinese Traditional ; Nerve Tissue Proteins ; metabolism ; Nestin ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism ; Stem Cells ; cytology

OBJECTIVETo determine cell proliferation and nestin expression in the ependyma of adult rat spinal cord after injury.

METHODSRat spinal cord injury models were established by aneurysm clip compression, and nestin expression and proliferation of ependymal cells at different times were shown with pathological and immuno-histochemical staining.

RESULTSEpendymal cells adjacent to the injured site demonstrated a dramatic increase in nestin expression 24 hours after compression. Proliferating cell nuclear antigen was positive, and significant proliferation was observed after 7 days. Nestin expression was down regulated as time went by.

CONCLUSIONNormally quiescent mature ependymal cells appear to revert to an embryonic state in response to spinal cord injury.

Animals ; Cell Division ; Ependyma ; cytology ; metabolism ; Immunohistochemistry ; Intermediate Filament Proteins ; biosynthesis ; Male ; Nerve Tissue Proteins ; Nestin ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; metabolism ; pathology

OBJECTIVETo identify the type of the intermediate filament (IF) protein of Angiostrongylus cantonensis and analyze its tissue localization.

METHODSRecombinant pET-IF of antigen IF was expressed in E.coli with IPTG induction, and the expression products were purified by His.Bind column and identified for determining the type of the IF protein by Western blotting. Anti-IF antibody was prepared by multi-spot subcutaneous injection into mouse and used to detect the tissue slices of A. cantonensis by immunohistochemical analysis.

RESULTSThe antigen IF were correctly expressed and purified, and identified as a keratin located in the intestine wall and cytoplusma.

CONCLUSIONThe antigen IF is distributed in the intestine wall of A. cantonensis.

Angiostrongylus cantonensis ; cytology ; metabolism ; Animals ; Cell Nucleus ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Intermediate Filament Proteins ; classification ; genetics ; isolation & purification ; metabolism ; Protein Transport

BACKGROUNDMany researchers suggest that adult mammalian central nervous system (CNS) is incapable of completing self-repair or regeneration. And there are accumulating lines of evidence which suggest that endogenous neural stem cells (NSCs) are activated in many pathological conditions, including stroke in the past decades, which might partly account for rehabilitation afterwards. In this study, we investigated whether there was endogenous neural stem cell activation in intracerebral hemorrhagic (ICH) rat brains.

METHODSAfter ICH induction by stereotactical injection of collagenase type VII into globus pallidus, 5-Bromo-2 Deoxyuridine (BrdU) was administered intraperitoneally to label newborn cells. Immunohistochemical method was used to detect Nestin, a marker for neural stem cells, and BrdU.

RESULTSNestin-positive or BrdU-Labeled cells were predominantly located at 2 sites: basal ganglion around hemotoma, ependyma and nearby subventricular zone (SVZ). No positive cells for the 2 markers were found in the 2 sites of normal control group and sham group, as well as in non-leisioned parenchyma, both hippocampi and olfactory bulbs in the 4 groups. Nestin+ cells presented 4 types of morphology, and BrdU+ nucleus were polymorphologic. Positive cell counting around hemotoma showed that at day 2, Nestin+ cells were seen around hemotoma in model group, the number of which increased at day 4, day 7 (P <0.01), peaked at day 14 (P <0.05), and reduced significantly by day 28 (P <0.01).

CONCLUSIONEndogenous neural stem cells were activated in experimental intracerebral hemorrhagic rat brains.

Animals ; Brain ; pathology ; Bromodeoxyuridine ; metabolism ; Cerebral Hemorrhage ; pathology ; Intermediate Filament Proteins ; analysis ; Male ; Nerve Tissue Proteins ; analysis ; Nestin ; Neurons ; pathology ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; pathology

OBJECTIVETo investigate the differentiative capability of adult human bone marrow mesenchymal stem cells (BMSCs) into Schwann-like cells.

METHODSBone marrows were aspirated from healthy donors and mononuclear cells were separated by Percoll lymphocytes separation liquid (1.073 g/ml) with centrifugation, cells were cultured in DMEM/F12 (1:1) medium containing 10% fetal bovine serum (FBS), 20 ng/ml epidermal growth factor (EGF) and 20 ng/ml basic fibroblast growth factor (bFGF). Cells of passage 1 were identified with immunocytochemistry.

RESULTSMononuclear cells separated by Percoll's were passaged 10 times by trypsin/ethylenediaminetetraacetic acid (EDTA) digestion in 40 days, and BMSCs increased about 6x10(7) times in this short period. Immunohistochemistry identified that BMSCs were CD34- and CD31-, but they expressed neuron specific enolase; 0.01%-0.02% of total cells expressed nestin, the marker for neural progenitor cells; 40%-50% cells stained heavily by neurofilament 200; and no glial fibrillary acidic protein (GFAP) positive cells were identified; S100 expression was detected among 0.1%-0.2% cells.

CONCLUSIONSBone marrow contains the stem cells with the ability of differentiating into Schwann-like cells, which may represent an alternative stem cell sources for neural transplantation.

Adult ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; physiology ; Cell Proliferation ; Humans ; Immunohistochemistry ; Intermediate Filament Proteins ; metabolism ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Nerve Tissue Proteins ; metabolism ; Nestin ; Neurofilament Proteins ; metabolism ; Phosphopyruvate Hydratase ; metabolism ; S100 Proteins ; metabolism ; Schwann Cells ; cytology

Astrocytoma ; pathology ; Brain Neoplasms ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Ganglioneuroma ; metabolism ; pathology ; surgery ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Intermediate Filament Proteins ; metabolism ; Male ; Nerve Tissue Proteins ; metabolism ; Nestin ; Neurons ; pathology ; Oligodendroglioma ; pathology ; Synaptophysin ; metabolism ; Vimentin ; metabolism ; Young Adult

OBJECTIVETo investigate the expressions of nestin and vascular endothehal growth factor (VEGF) mRNAs in rat brain tissue after cerebral ischemia-reperfusion injury and their changes in response to Tongxinluo (a traditional Chinese herbal preparation) treatment.

METHODSCerebral ischemia was induced in rats by temporary middle cerebral artery occlusion (MCAO) followed by treatment with Tongxinluo at high and low doses. On days 3, 7, 14 and 21 after MCAO, nestin and VEGF mRNA expressions in the ependyma, subventricular zone (SVZ), and hippocampal subdentate gyrus zone (HDG) in the ischemic hemisphere were quantitatively analyzed using immunohistochemistry and RT-PCR.

RESULTSCompared with the sham-operated group, the rats with cerebral ischemia-reperfusion injury showed significantly increased nestin-positive neurons and VEGF mRNA expression in the SVZ and HDG 7, 14 and 21 days after MCAO (P<0.05). Treatment with Tongxinluo, especially at high doses, significantly increased the number of nestin-positive neurons and VEGF mRNA expression in the rats 7, 14 and 21 days after MCAO (P<0.05).

CONCLUSIONFocal cerebral ischemia in rats results in rapid response and proliferation of neural stem cells in the SVZ and HDG in the ischemic hemisphere possibly by increasing VEGF mRNA expression in the adjacent tissues around the ischemic focus. Tongxinluo may enhance the differentiation and proliferation capacity of the neural stem cells after MCAO by inducing the expression of VEGF mRNA.

Animals ; Brain Ischemia ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Intermediate Filament Proteins ; genetics ; metabolism ; Male ; Nerve Tissue Proteins ; genetics ; metabolism ; Nestin ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism