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2008, Volume 39 Issue 1
06 February 2008 Previous Issue    Next Issue
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THE EXPRESSION OF CYCLIN DEPENDENT KINASE 11 FOLLOWING TRANSECTED SPINAL CORD INJURY IN RATS 2008, 39 (1):  1-6.  doi: Abstract ( )   Objective To explore the expression and distribution of cyclin dependent kinase 11 (CDK11) after transected spinal cord injury (tSCI). Methods tSCI was performed at T9 in adult SpragueDawley (SD) rats. The rats were sacrificed 1day, 3days, 5days, 7days, 14days after operation respectively. Western blotting was used to detect the changes of CDK11 protein expression after tSCI,and the distribution and localization of CDK11 in spinal cord were investigated by immunohistochemical staining and immunofluorescence double staining. Results Western blotting analysis showed the expressions of CDK11SUP>p58/SUP> and CDK11SUP>p110/SUP>, which were the two major isoforms of CDK11, increased at first and then had a decrease tendency in both broken ends of the transected spinal cord. CDK11SUP>p58/SUP> was enhanced on the 3rd day and remained high level until the 7th day; while CDK11SUP>p110/SUP> was also increased, on the 3rd day but went down rapidly after the 5th day. Immunohistochemistry staining was indicated that CDK11 was distributed evenly in the normal spinal cord. On the 3rd day, the staining was elevated obviously in both white and grey matter. Immunofluorescence double staining suggested that CDK11 was colocalized with neuronal neclei (NeuN), cyclic nucleotide 3′phosphohydrolase （CNPase） and glial fibrillary acidic protein (GFAP).Conclusion The expression of CDK11 undergoes spatiotemporal changes after tSCI. These alterations may be Related Articles | Metrics

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EFFECTS OF NERVE REGENERATION FACTOR ON THE NEURITE GROWTH OF RAT HIPPOCAMPAL NEURONS 2008, 39 (1):  7-11.  doi: Abstract ( )   Objective To determine the effects of nerve regeneration factor (NRF) on neurite growth in cultured hippocampal neurons of rats. Methods After different periods (6h, 12h, 24h) of incubation by different concentrations (0.25mg/L, 0.5mg/L, 1.0mg/L) of NRF, the hippocampal neurons were observed and photographed by phase contrast microscopy, and the neurite length was analyzed using the Scion software. Realtime fluorescence quantitative RT-PCR was performed to examine the mRNA levels of growth associated protein 43 (GAP-43) after different incubation periods with different concentrations of NRF. Fluorescent immunocytochemistry and Western blotting were performed to further examine the protein levels of GAP-43 in cultured hippocampal neurons after 24h incubation with different concentrations of NRF. Results NRF promoted the neurite growth of cultured hippocampal neurons with the most obvious mophological changes appearing upon treatment with NRF at 1mg/L for 24 hours. Realtime fluorescence quantitative RT-PCR showed that the mRNA level of GAP-43 in cultured hippocampal neurons was upregulated by NRF in a dose and timedependent manner, with the best concentration of 1mg/L. Meanwhile, fluorescent immunocytochemistry and Western blotting showed that the protein level of GAP-43 was upregulated by NRF with the best concentration of 1mg/L. Conclusion NRF could promote the neurite growth and modulate the expression of GAP-43 in cultured hippocampal Related Articles | Metrics

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RESEARCH ON BONE MARROW MESENCHYMAL STEM CELLS MODIFIED BY NT-3 GENE AND PRETREATED WITH RETINOIC ACID TO DIFFERENTIATE INTO NEURONLIKE CELLS IN THE INJURED SITE OF SPINAL CORD 2008, 39 (1):  12-17.  doi: Abstract ( )   Objective To explore the potential of bone marrow mesenchymal stem cells (MSCs) to be modified by neurotrophin3 (NT-3) gene and pretreated with retinoic acid (RA) to differentiate into neuronlike cells in the transplanted site of the spinal cord injury. Methods MSCs, RAinduced MSCs, LacZ gene modified MSCs, NT-3 gene modified MSCs, and MSCs both modified by NT-3 gene and pretreated with RA were immediately transplanted respectively into the completely transected site (TSUB>10/SUB> spinal segment) of spinal cord. On the 67th day after the operation, the spinal cord segment was removed and frozenly sectioned. The differentiation potential of MSCs was examined by immunofluorescence histochemistry and the percentage was calculated of neuronlike cells that were differentiated from MSCs among all the transplanted cells groups. Results Transplanted MSCs could differentiate into neural stem cells (nestinpositive), neuroglial cells (GFAPpositive) and neuronlike cells (NF and MAP2positive) in the injured spinal cord. Some of them also differentiated into the neuronlike cells which contained some neurotransmitters (ChAT and 5-HT positive) or had the potential to form a synapse (PSD95_positive). The percentage of neuronlike cells differentiated from MSCs modified by NT-3 gene and pretreated with RA was the highest among all the transp Related Articles | Metrics

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EFFECT OF MONOCHROMATIC LIGHT ON CELL DENSITY AND CELL SIZE OF RETINAL GANGLION CELLS IN CHICKS 2008, 39 (1):  18-22.  doi: Abstract ( )   Objective To study the effect of monochromatic light on the distribution of retinal ganglion cells in chicks. Methods Sixty One-day-old male broilers were reared under four light treatments, red (660nm), green (560nm), blue (480nm) and white (400-760nm) by using LED (light-emitting diodes) as light sources until the 49th day (n=15). Light intensity was 15 lux at the height of birds’ heads and scheduled for 23hours of light and 1hour of darkness during the entire experiment. The retinas were stained by Nisslstaining and DiIlabeling. The retinal area, cell size and density in ganglion cell layer were estimated by image analysis. Results The retinal area and RGCs’ number in the blue and green light groups were higher than that in the red and white light groups (10.35% and 17.07%, P<0.05). The average cell density of RGCs in the green light group was higher than that in other groups (14.61%, P<0.05). The cell density of RGCs in the green light group was the highest at retinal central area and was higher than that in the blue light group (28.91%, P<0.05). The ratio of CA/NP in the green light group was higher than that in the others (29.71%, P<0.05). The cell size of RGCs in the blue light group was the smallest in central area and the highest in periphery (TP and NP). The ratio of TP/CA in the blue light group was higher than that in the others (26.65%, P<0.05) and there was a significant difference between the others (P<0.05). Conclusion Blue and green lights increased retinal area and RGCs’ number. The decreased amplitude of RGCs’ density gradient in the green light group and the increased amplitude of RGCs’ size gra Related Articles | Metrics

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THE INFLUENCE OF CARBON TETRACHLORIDE UPON THE EXPRESSION OF LOWAFFINITY NEURAL GROWTH FACTOR RECEPTORp75 IN HIPOCAMPAL CA1 AREA NEURONS 2008, 39 (1):  23-26.  doi: Abstract ( )   Objective This study aimed to research the influence of CCl-4 on the expression of p75 and apoptosis in hippocampal CA1 area neurons. Methods Healthy adult male Wistar rats were randomly divided into control group and CCl-4 groups. The control group was subcutaneously injected olive oil solution with 012ml/100g rat weight on every Monday or/and Thursday, the CCl-4 groups were subcutaneously injected 60% carbon tetrachloride olive oily solution with 0.3ml/100g rat weight on every Monday or/and Thursday for one time(one day group), two times(one week group), four times(two_week group) respectively. When the experiment ended, all the rats were killed. Brains were taken out and separately slivered into two parts along median sagittal plane. The right parts were embedded in paraffin wax and sliced, the morphological changes of hippocampal CA1 area neurons were observed by Nissl staining. The expressions of p75 and Bax in hippocampal CA1 area neurons were detected by immunohistochemical staining and Western blotting analysis. The apoptosis of hippocampal CA1 area neurons was observed by TUNEL staining. Results The color of Nissl staining was deeper,the number was more and the structure was more regular than that of the CCl-4 groups, and there were more Nissl’s bodies in the control group; while in the CCl-4 groups, the staining was lighter and the structure was less regular than that of the control group, some pyramidal cells shrunk, their nuclear dwindled and presented triangles, and there were less or no Nissl’s bodies. There were a few p75 and Bax positive cells in the control group, but p75 and Bax positive cells increased obviously in every CCl-4 group; and increased most obviously after injecting CCl-4 for two times. In Western blotting, the expression of p75 and Bax was the same as the results of immunohistochemical staining. There was no TUNEL positive cell in the control group, but more brown nucleus_stained TUNEL positive cells could be observed in the CCl-4 groups,and the one week CCl-4 group presented the most TUNEL positive cells. Conclusion After the injection of carbon tetrachloride, the expression of p75 increased evidently in hippocampal CA1 area neurons, the expression of Bax enhanced correspondently, apoptotic neurons also increas Related Articles | Metrics

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THE MORPHOLOGICAL ANALYSIS OF BRAIN TISSUE FROM TRANSGENIC MOUSE WITH EXPRESSED GREEN FLUORESCENT PROTEIN IN NERVOUS TISSUES 2008, 39 (1):  27-30.  doi: Abstract ( )   Objective To establish transgenic mice with nervous tissues specifically expressing green fluorescent protein and to make an animal model for the study of nervous system by fluorescent imaging. Methods The transgenic plasmid was constructed by inserting the EGFP gene under the control of PDGF Bchain promoter and transgenic C57BL/6J mice were established by microinjection method. The transgenic mice were genotyped by PCR. The total of 48 pairs of freezing sections of transgenic mice were prepared in the sagittal plane, and every two adjacent sections were observed under microscope and compared with HE staining and fluorescent imaging. Results One transgenic line with high levels of green fluorescent protein expression was detected from eight transgenic lines. Green fluorescent protein expression was observed in the tissues of cerebrum, hippocampus, thalamencephalon, cerebellum and brain stem. Conclusion A transgenic mouse line was established with nervous tissues specifically expressing green fhcore sunt protein. The results suggested that it will be an available animal model for the study of nervous tissues by fluorescent imaging. Related Articles | Metrics

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THE EXPRESSION OF BONE MORPHOGENETIC PROTEIN7 IN THE HYPOXIC AND ISCHEMIC CAUDALPUTAMEN CELLS OF RATS 2008, 39 (1):  31-34.  doi: Abstract ( )   Objective To study the expression of bone morphogenetic protein7(BMP_7) in the hypoxic and ischemia damage striatum tissues, and investigate the neuroprotective effect of endogenous BMP-7 in the injuried nerve tissues of rats. Methods Rat models of focal cerebral ischemia were made. Immunohistochemistry method was used to detect the expression of BMP-7 in the rat striatum 24 hours after ischemia/reperfusion and caudalputamen cells were cultured under hypoxia/reoxygenation. The absorbance of the positive products of the stained section was measured by computer image processing software, and statistics analysis was done. Results The expression of BMP-7 in the caudalputamen was higher and more extensive in the ischemic side than that in the nonischemic side 24 hours after ischemia/reperfusion, and there was a statistically significant difference between the average absorbance in the ischemic side and that in the nonischemic side. (P<0.01), while BMP-7 expression was not detected in the bilateral caudalputamen in the normal and sham control rat groups. In the primary cultural cells, the weak expression of BMP-7 was detected in the plasm of caudalputamen cells 24 hours after hypoxia/reoxygenation, while no BMP-7 positive expression was found in the normal control caudalputamen. Conclusion This experiment explained that hypoxicischemic can cause endogenous BMP-7 expression. The increased expression of BMP-7 ind Related Articles | Metrics

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THE INTERFERING EFFECT AND PROBABLE MECHANISM OF NEUREGULIN-1β ON CEREBRAL ISCHEMIA REPERFUSION INJURY IN MICE 2008, 39 (1):  35-39.  doi: Abstract ( )   Objective To study the neuroprotective effects of neuregulin-1β (NRG-1β) on the nervous behavioral function, cerebral infarction volume, brain water content(BWC), neuronal apoptosis and aquaporin4(AQP-4) expression in astrocytes after cerebral ischemic reperfusion and the related mechanism in mice. Methods Intraluminal thread methods were applied to establish the middle cerebral artery occlusion reperfusion models in the mice. Neuregulin-1β (2μg / kg) was injected into the internal carotid artery for treatment. The nervous behavioral function was evaluated with Bederson’s test. The cerebral infarction volume was observed with tetrazolium chloride staining. The BWC was measured by drywet weight comparing. The apoptosis positive cells were counted by immunofluorescence assay. The expression of AQP-4 was determined by immunohistochemical assay. Results Nervous behavioral malfunction appeared in all the mice with left middle cerebral artery occlusion and/or reperfusion. The infarction focus showed in the ischemic hemisphere after the injury. The BWC, the number of neuronal apoptosis cells and AQP-4 expression in astrocytes were higher than those in the sham group. In the NRG-1β treatment group, the nervous behavioral function was improved 24 hours after ischemia, the number of apoptosis positive cells reduced and the infarction volume decreased significantly compared with the control group(P<0.05). The BWC and AQP-4 expression in astrocytes showed no significant difference compared with the control group(P>0.05). In the groups of reperfusion for 22,46 and 70 hours,the five indexes mentioned above were significantly different from those in the corresponding control groups(P<0.05). Conclusion NRG-1β may reduce cerebral edema and infarction volume and improve the nervous behavioral functio Related Articles | Metrics

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A STUDY ON THE EFFECT OF OLFACTORY ENSHEATHING CELLS ON THE PROLIFERATION OF NEURAL STEM CELLS- 2008, 39 (1):  40-44.  doi: Abstract ( )   Objective To research the effects of olfactory ensheathing cells (OECs) on the proliferation of neural stem cells (NSCs). Methods OECs and NSCs were separated and cultured in vitro. The proliferation of NSCs was observed by means of co-culture and twochamber shared medium culture with OECs. For investigating the mechanism of the proliferation of NSCs, the cytokines and their receptors of OECs were tested by immunocytochemistry and RT-PCR.Results After 4 days in co-culture and twochamber shared medium culture, the population of NSCs were increased significantly (P<0.05), and some of them were differentiated. The result of RT-PCR showed that OECs could express mRNA of β-NGF, BDNF, NT-3, PDGFB, trkA and trkB, but not NT-4 Moreover, positive staining of NGF, BDNF and p75 was observed in the cytoplasm of OECs by immunocytochemistry. Conclusions OECs can express or secrete a l Related Articles | Metrics

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THE EFFECTS OF CYTOKINES ON THE MIGRATION AND MOBILIZATION OF LYMPHATIC ENDOTHELIAL PROGENITOR CELLS 2008, 39 (1):  45-49.  doi: Abstract ( )   Objective To investigate the effects of VEGF-C，SDF-1，MCP-1 and G-CSF on migration and mobilization of CD34+/CD133+/VEGFR-3+ lymphatic endothelial progenitor cells (LEPCs). Methods Mononuclear cells were isolated from canine peripheral blood by noncontinuous density centrifugation with Percoll solution. VEGFR-3+ cells were sorted with flow cytometxy. Expression of specific marks CD34 and CD133 on VEGFR-3+ cells were observed under a confocal laser scanning microscope. The effects of VEGF-C，SDF-1，MCP-1 and G-CSF on chemotatic migration of LEPCs were examined by transmigration assay. The morphological characteristics of the transmigrated cells were observed under scanning electron microscope. VEGF-C, SDF-1, MCP1 and G-CSF were administrated to rats by subcutaneous injection.The number of LEPCs in peripheral blood was analyzed with flow cytometry and white blood cells were counted with blood cell analyzer. Results The percentage of the migrated cells through the membrane was greater in the VEGF-C, SDF-1, MCP-1 and G-CSF groups than that in the control group. After the cells were treated with blocking antibodies of VEGFR-3 and CXCR4, the percentage of the migrated cells decreased obviously. After the injections of VEGF-C, SDF-1, MCP-1 and G-CSF, LEPCs in peripheral blood of rats increased obviously, while significant increase of white blood cells was observed. Conclusion VEGF-C, SDF-1, MCP-1 and G-CSF play a role in the chemotatic migration Related Articles | Metrics

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THE EXPRESSION OF Oct-4 and hTERT ON HUMAN PRIMORDIAL GERM CELLS AT DIFFERENT PERIODS OF DEVELOPMENT 2008, 39 (1):  50-54.  doi: Abstract ( )   Objective To analyse the expression levels of Oct-4 and human telomerase reverse transcriptase(hTERT) in human primordial germ cells (PGCs) which were isolated at different fetus ages. Methods The expressions of both Oct-4 and hTERT were detected by RT-PCR. The gonadal ridges of human embryos were embedded with paraffine and sliced, and stained with HE. The morphology of the human primordial germ cells was observed. The gonadal ridges were analyzed for the expression of Oct-4 and hTERT through immunohistochemistry. Results There were a few PGCs in the gonadal ridges in the sixth to the seventh week of gestation, and both Oct-4 and hTERT were expressed in the PGCs. The PGCs with Oct-4 and hTERT positive staining increased in number in week 8 to 12 of gestation (P<0.05). The PGCs decreased in number and the Oct4 immunostaining intensity in PGCs declined, but the hTERT was still expressed in PGCs in the thirteenth week of gestation. Conclusion The immunostaining intensity of Oct-4 varies with different stages of development and is the highest during week 8 through 12 There was no obviously change on the expression of hTERT. Related Articles | Metrics

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THE STUDY OF IMMUNOGENICITY OF NONHEMATOPOIETIC STEM CELLS FROM UMBILICAL CORD BLOOD 2008, 39 (1):  55-59.  doi: Abstract ( )   Objective To explore the immunogenicity of nonhematopoietic stem cells from human umbilical cord blood (HUCB). Methods We detected HLA-ABC, HLA-DR of HUCBnHSCs and HUCBnHSCs treated of IFN-γ for 3 days by FACS. Then CB-nHSCs, passage 1 CB-nHSCs, differentiated CB-nHSCs, CB-nHSCs treated with IFN-γ, human neuroblastoma cell line SH-SY5Y were co-cultivated respectively with spleen lymphocytes to observe proliferation of these lymphocytes.Results The HUCBnHSCs were positive for HLA-ABC, while few cells were positive for HLA-DR. The treatment of IFN-γ didn’t increase the expressions of HLA-ABC and HLA-DR. In mixed lymphocyte culture, they all failed to stimulate proliferation of lymphocytes except for the SH_ Related Articles | Metrics

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CULTURE AND BIOLOGICAL PROPERTIES OF ADULT ESOPHAGEAL EPITHELIAL CELLS IN VITRO 2008, 39 (1):  60-63.  doi: Abstract ( )   Objective To culture normal adult esophageal epithelial cells and establish a normal esophageal epithelial cell line to provide experimental materials for further studies in vitro. Methods Normal esophageal epithelium samples were obtained in sterile phosphate buffered saline supplemented with antibiotics from the normal part of the esophagus of a patient with esophageal carcinoma. Tissue was cleaned of all connective tissues and muscles and rinsed in phosphate buffered saline 5_6 times. The mucosal layer was minced into small pieces and dissociated into a single cell suspension by 025% Dispase and 025% trypsin/002%EDTA. Cells were grown in keratinocyte serumfree media. The cultured cells were identified through their morphological characteristics and immunohistochemical staining. The proliferative capacity of the cultured cells was also examined. Results The cultured cells showed microscopic features of epithelial cells and were positive in keratin and epithelial membrane antigen staining. Eight days after primary culture, the cells displayed a cobblestone morphology reaching 80%_90% confluency and were passaged successfully with 025% trypsin/002%EDTA. The cell cycle analysis showed about 7760% of the cultured cells was in G0/G1 phase and the others in S/G2/M phase. Conclusion\ The culture methods and techniques used in the exp Related Articles | Metrics

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EFFECT AND POSSIBLE MECHANISMS OF PANAXYDOL ON NTFs EXPRESSION IN SCHWANN CELLS IN VITRO 2008, 39 (1):  64-69.  doi: Abstract ( )   Objective To study the effects of panaxydol(PND) upon neurotrophic factors (NTFs) expression in Schwann cell, and to further reveal the mechanisms that underlie these effects. Methods Schwann cells were treated with PND of different concentrations in the culture medium. NTFs expression was examined through immunoiistochemistry assay. The intracellular cAMP level was assessed by radioimmunologic assay. Results There was an increase in NGF expression after PND treatment within the range of 2.5-200μmol/L (P<0.05) and an increase in brain derived neurotrophic factor(BDNF) expression after PND treatment within the range of 5.0-20.0μmol/L (P<0.05). Both NGF and BDNF expressions peaked in 10μmol/L PND group with P<0.01, which was considered to be statistically significant; Intracellular cAMP increased as well after PND treatment within the range of 2.5-20.0μmol/L (P<0.05) and the intracellular cAMP level also peaked in the 10μmol/L PND group (P<0.01). Conclusion PND treatment of Schwann cell enhances NGF and BDNF expressions in vitro. In addition, an increase in the intracellular cAMP level was observed after PND treatment and that correlates with NGF and BDNF expression Related Articles | Metrics

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THE EXPRESSIONS AND LOCALIZATIONS OF B7-H1 IN DIFFERENT GASTRIC CANCER CELL LINES 2008, 39 (1):  70-73.  doi: Abstract ( )   Objective B7-H1 is a recently identified costimulatory molecule expressed on APCs and can inhibit the activation and function of T lymphocytes. This research is to study if B7-H1 has a relationship with carcinogenesis and MDR of gastric carcinoma by examining the expressions of B7-H1 in human gastric carcinoma cell lines SGC7901/VCR,SGC7901,BGC823 and in the permanent gastric epithelial cell line GES1. Methods Cells were cultured in RPMI1640 supplemented with 10% fetal calf serum. The B7-H1 mRNA was detected by RT-PCR and the protein of B7-H1 was detected by cytoimmunochemistry staining, cytoimmunofluorescence staining and flow cytometric analysis. The expression levels of B7-H1 mRNA and the protein of B7-H1 in the above mentioned four kinds of cell lines were compared. Results All the four cell lines express B7-H1 mRNA at different levels. And B7-H1 expression level increased in the order of from GES-1 to SGC7901/VCR cell line, BGC-823 and then to SGC7901 cell line. The protein of B7-H1 was found to be distributed in the cell membrane and in a little cytoplasm in all the four cell lines through techniques of cytoimmunochemistry staining and cytoimmunochemistry staining. Further more with the result of flow cytometric analysis, the protein expression level of B7-H1 in all these cell lines was found to be consistent with B7-H1 transcript level expression.Conclusion The results suggest that B7-H1 expression on gastric epithelial cells may promote the growth of gastric carcinoma cells and the expression of MDR gene through inh Related Articles | Metrics

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A STUDY ON THE EVOLUTION OF FLAMINGO CADHERIN SUBFAMILY 2008, 39 (1):  74-78.  doi: Abstract ( )   Objective To perform phylogenetic study on the flamingo cadherin subfamily and explore the evolutionary differences between the flamingo cadherins of vertebrates and those of invertebrate. Methods Phylogenetic trees were established, the protein domains were compareds and the gene structure analyses was conducted. Results The flamingo cadherin subfamily was originated from eumetazoa. In vertebrates,two duplication events occurred and resulted in the three paralogs. Intron insertions were found in the vertebrate flamingo protein domains, and two highly conserved motifs were also showed in the vertebrate flamingo cytoplasm. Both of them can be viewed as the evolutionary characteristics of vertebrate flamingo cytoplasm. Conclusion There are significant evolutionary differences between the flamingo cadheins of vertebrates and those of invertebrates. Together with the positioning information that the Celsr1 gene,one of the mouse flamingo paralogs,is located in the quantiative trait loci(QTL) which is related to mice spinal cord development, Celsr1 can be considered as a candidate gene of mice spinal cord development for further studies. Related Articles | Metrics

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PROLIFERATIVE EFFECT OF INSULINLIKE GROWTH FACTOR-1 ON MUSCLEDERIVED STEM CELLS 2008, 39 (1):  79-82.  doi: Abstract ( )   Objective To explore the effect of insulinlike growth factor1 (IGF-1) on the growth of musclederived stem cells (MDSCs). Methods MDSCs were isolated from the hindlimb muscle of new born mice through serial preplates. DMEM with 2% fetal bovine serum was used to induce MDSCs to differentiate into skeletal muscle lineage. The expression of stem cell marker Sca-1 or skeletal muscle cell marker α-sarcomeric actin was examined by immunocytochemistry SP method. MTT colorimetric microassay was conducted to determine the effect of IGF-1 on MDSCs proliferation. The relationship between functioning and culture time and the relationship between functioning and IGF-1 concentration were studied. Results MDSCs were successfully isolated from the hindlimb of neonatal mice. Over 90% of the MDSCs showed Sca-1 positive immunoreactivity. MDSCs could give rise to α-sarcomeric actin positive myotubes in differentiation cultures. The proliferative effect of IGF-1 on MDSCs increased gradu ally with prolonged culture time. The elevation of IGF-1 concentration also increased the proliferative effect until it came to a saturation point.Conclusion Related Articles | Metrics

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STUDY OF THE EXPRESSION OF GERM CELL DIFFERENTIATION ASSOCIATED GENES ON HUMAN EMBRYONIC STEM CELLS 2008, 39 (1):  83-87.  doi: Abstract ( )   Objective To determine whether human embryonic stem cell (hES) line H1 can express the germ cell differentiation associated genes in vitro. Methods The characteristics of H1 was analyzed with the expression of alkaline phosphatase, immunologic markers and pluripotency markers. Giemsa staining was used to indicate the karyotype of H1. The expression of germ cell differentiation associated genes on H1 was detected by RT-PCR. The normal human testis and human embryonic fibroblast cell line HFF-1 served as the positive and negative controls respectively. Results H1 remained the normal karyotypes 46 and XY and maintained the proliferation in an undifferentiated state. H1 expressed high levels of alkaline phosphatase(AP). However, unlike mouse embryonic stem cells (mES), H1 did not express SSEA-1(stagespecific embryonic antigen1), but expressed the SSEA4, TRA-160, TRA-181 and OCT4. In addition, H1 expressed OCT4 and NANOG, which presented the specific gene of pluripotency and selfrenewal. Premeiotic germ cellspecific genes were expressed in H1, including STELLAR (STELLArelated)，DAZL(deleted in azoospermialike)，PUM1 (PUMILIO 1)，PUM2 (PUMILIO 2)，FRAGILIS and GDF3(growth and differentiation factor 3). H1 did not express the germ cellspecific genes of meiotic and postmeiotic differentiation, including the meiosis initiating gene STRA8(stimulated by retinoic acid gene 8), meiosis specific gene SCP1(synaptonemal complex protein 1), germ cell mature differentiation gene VASA, PGCs (primordial germ cells) specific gene BLIMP-1 Related Articles | Metrics

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THE EFFECT OF INTRAMYOCARDIAL TRANSPLANTATION OF EMBRYONIC STEM CELLS COMBINED WITH BASIC FIBROBLAST GROWTH FACTOR INTO MYOCARDIAL INFARCTION MODELS ON THE CARDIAC FUNCTION IN POSTINFARCTED RATS 2008, 39 (1):  88-92.  doi: Abstract ( )   Objective To investigate the feasibility of recovering the cardiac structure and improving the cardiac function after the transplantation of embryonic stem cellsD3 (ES-D3) combined with basic fibroblast growth factor (b-FGF) into the acute myocardial infarction (AMI). Methods Forty Wistar rats were used and randomly divided into 5 groups equally, including the normal control group (group 1), the myocardial infarction without treatment group (group 2), the medium injected group (group 3), the ES-D3 transplanted group (group 4),the combination of ES-D3 and bFGF transplanted group (group 5).One week after the acute myocardial infarction (AMI), rats were injected with the in vitro differentiated ES-D3 labeled by BrdU. Then the cardiac function and histology were measured in week 4 after the transplantation. Results ES-D3 could differentiate into cardiomyocytes in vitro, and could survive steadily for 4 weeks in vivo after the transplantation into AMI’s model. There were no significant difference between group 2 and group 3 in cardiac function（P>0.05）. Compared with group 2, the infarction size and left ventricular weight(LVW) were reduced significantly（P<0.01）in group 4 and group 5， and capillary density was increased significantly（P<0.01）. The functions of the left ventricle were remarkably improved. Conclusion The t Related Articles | Metrics

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THE STUDY ON MECHANISM OF MINOXIDIL’S PROMOTING EFFECT ON CULTURED ADULT HAIR FOLLICLES 2008, 39 (1):  93-97.  doi: Abstract ( )   Objective To investigate the mechanism of minoxidil’s promoting effect on cultured adult hair follicles. Methods Different concentrations of minoxidil was added on adult hair follicles cultured in serumfree medium. Hair growth was measured at different time points. The amount of collagen I, metalloproteinases1(MMP-1), transforming growth factor β1(TGF β1), integrin β1 and K19 were tested by histochemistry and immunohistochemistry. Results Cultured hair follicles grew fast during the first three days, and then grew slow afterwards. 1mg/L of minoxidil increased the growth of hair follicles most effectively among the concentrations from 0.1mg/L to 100mg/L. The appearance of catagen appeared later in minoxidil group than control. The expression of collagen I was stronger in minoxidil group than in control; the expression of MMP-1 and TGFβ1 were less. There was no significant difference of the expression of integrin β1 and K19 between experiment and control groups.Conclusion Minoxidil can promote the g Related Articles | Metrics

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THE DISTRIBUTION AND EXPRESSION OF IL-18 IN RATUTERUS DURING PERIIMPLANTATION 2008, 39 (1):  98-101.  doi: Abstract ( )   Objective To study the distribution and expression of interleukin-18(IL-18) in rat uterus during periimplantation and understand possible physiological regulatory roles of endogenous IL-18 in embryo implantation. Methods Immunohistochemical technique and image analysis were used. Results IL-18 protein could be detected in all the rat uteri. The positive cells were mainly distributed in the uterine endometrial or decidual luminal epithelium, the glandular epithelium,and the stroma or decidual cells; a weak expression was also found in the myometrium and some endothelial cells. Compared with the estrus group, the expression of IL-18 in pregnant rats distinctly increased (P<0.05）, and with the developing of pregnancy, the level of IL18 in pregnant rats was gradually enhanced. Moreover, the expression of IL-18 during postimplantation period was markedly higher than that during preimplantation period (P<0.05）.Conclusion IL-18 may play a role in pregnancy and regulate embryo implantation.It m Related Articles | Metrics

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THE REGULATION OF EXOGENOUS POLYAMINES ON ODC PROTEIN LEVEL IN THE EARLY REGENERATING LIVER OF RATS 2008, 39 (1):  102-105.  doi: Abstract ( )   Objective To study the role of polyamines (putrescine, spermidine and spermine) in regulating ornithine decarboxylase (ODC) protein levels in the early regenerating liver of rats. Methods Exogenous polyamines (dissolved in 0.9% NaCl) were administered subcutaneously to adult male SD rats (180～220g). The ODC protein levels in the regenerating liver induced by partial hepatectomy (PH) were measured by Western blotting analysis. Results The treatment with high doses of putrescine ( 20 mg/kg body weight), spermidine (0.15 mg/kg body weight) or spermine ( 6 mg/kg body weight) reduced the ODC protein level compared with the control group within 12 hours after PH. However,the treatment with low doses of putrescine (0.02 mg/kg), spermidine (0.03 mg/kg) or spermine (0.06 mg/kg) resulted in a rapid increase in the protein level in the 4th hour after PH, which was 15.1%, 29.5% or 30.3% higher than that of control rats respectively.Conclusion Polyamines with certain doses play a role in the feedback regulation of ODC protein levels in the early regenerating liver, where spermidine and spermine play a greater role than putrescine dose. Related Articles | Metrics

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THE EXPRESSIONS OF ANNEXIN 5 mRNA AND BAX mRNA WITH FQ-RT-PCR IN THE RAT TESTIS WITH SUBMANDIBULAR GLAND REMOVED 2008, 39 (1):  106-110.  doi: Abstract ( )   Objective To detect the expressions of annexin 5 mRNA and Bax mRNA in the rat testis after the submandibular gland was removed. Methods On days 14, 28 and 42 after the operation, the expression levels of annexin 5 and Bax mRNA in the rat testis were examined by fluorogenic quantitative RT-PCR. Results Fluorogenic quantitative RT-PCR showed that there was a 38.5% and 55.3% increase in the expression of annexin 5 mRNA and a 70.6% and 80.5% significant increase in the expression of Bax mRNA (P<0.05, P<0.01) in the rat testis on days 14 and 28 respectively. There was no significant change in the expressions of annexin 5 and Bax mRNA on day 42 compared with the control groups.Conclusion The removal of submandibular gland can significantly increase the expressions of annexin 5 and Bax mRNA in the rat testis on days 14 and 28. The expressions of annexin 5 and Bax mRNA goes back to the normal level on day 42 Related Articles | Metrics

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A STUDY OF INCUBATION AND INITIAL IDENTIFICATION OF MURINE YOLK SAC MESENCHYMAL STEM CELLS 2008, 39 (1):  111-116.  doi: Abstract ( )   Objective To investigate the isolation, incubation and identification of murine yolk sac mesenchymal stem cells(YS_MSCs). Methods Murine yolk sacs were harvested on the 10th day of postcoitus microisolation.Yolk sac cells were obtained after the yolk sacs were digested by type Ⅰ collagenase of 0.1% for 1 hour. Adherent cells were cultured in DMEM containing 10% FBS and 1% SP,and passaged when they became subconfluent.The ultrastructure of YS_MSCs was observed under transmission electron microscope; the surface markers of YS_MSCs were detected by flow cytometry; the alkaline phosphatase(AKP) expression of YS_MSCs was tested; and cytochemical changes of YS_MSCs were observed with cytochemical detection. Adipogenic differentiation of YS_MSCs was induced by 1μmol/L dexamethasone and 10mg/L insulin, and oil red O was used for fat staining. Results YS_MSCs were obtained in vitro, and most of them were of spindle shape;under transmission electron microscope,there were microvilli on the surface of YS_MSCs,there were abundant mitochondria, rough endoplasmic reticulum and golgi complex in the endochylema. The cell nuclei of YS_MSCs were big and the shape of cell nucleus was irregular; the analysis of flow cytometry suggested that the primary and first generation of YS_MSCs were positive for CD44,CD105, and the expression of CD34 was small;Cytochemistry showed that YS_MSCs were positive for glycogen,and negative for SB and AKP; Adipogenic differentiation of YS_MSCs was induced by 1μmol/L dexamethasone and 10mg/L insulin;accumulation of lipid_rich vacuoles appeared in the cells, positive for oil red O staining.Conclusion The biological characteristics of murine yolk sac mesenchymal stem cells are similar to, and more primitive than, those of adult mesenchymal stem cells.It suggests that YS_MSCs may be used as an Related Articles | Metrics

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THE EFFECTS OF SODIUM SELENITE ON THE FUNCTION OF DENDRITIC CELLS FROM PERIPHERAL BLOOD OF CHRONIC HEPATITIS PATIENTS 2008, 39 (1):  117-120.  doi: Abstract ( )   Objective To explore the methods of improving the immune function of dendritic cells(DCs) from patients with chronic type B hepatitis in vitro. Methods Monocytes from type B hepatitis patients and healthy controls were incubated with GM-CSF and IL-4 and IFN-α to induce dendritic cells (DCs) generation. Then the nutrienconcentration sodium selenite was added for co-culture before the maturity of DCs. The samples were separated into three groups: hepatitisB patients (group A), hepatitisB patients with Se (group B), healthy control (group C). The phenotypes of DCs including CD80, CD86 were characterized by flow cytometry(FCM) and the response of allogeneic lymphocytes was detected by MTT assay. The concentration of cytokines in the supernatant such as IL-12 was tested by ELISA assay. Results After the nutrienconcentration Se was added for co-culture, the proliferation ability of isotype xenoma lymphocyte stimulated by DCs was obviously streng thened compared with group A, and the excretion level of IL-12 also increased (P<0.01).There was no significant difference between group B and group C.Conclusion Co-cultured with the nutrientconcentration Se in vitro, DCs could stimulate lymphocyte proliferation reaction effectively, improve the excretion of Related Articles | Metrics

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THE CULTURE OF MYOCARDIAL MICROVASCULAR ENDOTHELIAL CELLS OF SUCKLING RATS EM>IN VITRO/EM> 2008, 39 (1):  121-123.  doi: Abstract ( )   Objective To improve methods of culturing rat myocardial microvascular endothelial cells (RMMVECs) EM>in vitro/EM> and provide experimental data for studying the structures and functions of RMMVECs. Methods SD rats of 10_15 days old were selested. Explants cultivation of cardiac tissues was used to obtain the primary cells. In the subculturing, the cell cultures were purified by different speeds of attachment and digestion from contaminant cells combined with morphological observation. The purified cells were identified as endothelial cells by morphology and by positive immunocytochemistry for factor Ⅷrelated antigen, a marker for endothelial cells. Results The RMMVECs were cultured and purified successfully which grew in a polygonal or cobblestone pattern and were positive for the immunocytochemical staining of factor Ⅷrelated antigen. Conclusion The methods of culturing RMMVECs were improved, RMMVECs with high purity were o Related Articles | Metrics

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THE STUDY ON THE EXPRESSION OF BASIC FIBROBLAST GROWTH FACTOR AND TRANSFORMING GROWTH FACTORBETA IN THE SCLERA OF HUMAN FETUS 2008, 39 (1):  124-126.  doi: Abstract ( )   Objective To evaluate the existence of basic fibroblast growth factor(bFGF) and transforming growth factorbeta(TGF-β)in the sclera in different fetal ages by realtime polymerase chain reaction (PCR). Methods Forty human fetal eyes ranging in age from 12 to 20 pregnant weeks were carefully extracted. The sclera was carefully dissected. The expressions of bFGFand TGF-β were determined by realtime PCR, and the results were analyzed comparatively. Results bFGF was expressed in the highest amount at the 12th to 14th pregnant week and decreased thereafter, while the expression of TGF-β lowered at the 12th week and increased gradually.Conclusion bFGF and TGF-β express in the sclera of the human fetal Related Articles | Metrics

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THE 3D VISUALIZATION OF THE PULMONARY VENOUS SYSTEM OF CHINESE PEOPLE 2008, 39 (1):  127-130.  doi: Abstract ( )   Objective To build a digitized 3D model of the pulmonary venous system of Chinese people for the purpose of providing anatomic information of the pulmonary venous system for anatomic teaching, imaging diagnosis and surgical operation of pulmonary diseases. Methods The primal images from the first Chinese visible human data set of chest were converted into image sequence only cantaining the sections of pulmonary veins after manual registration and image segmentation. Then these images were imported into 3DMed software to build a 3D model of the pulmonary venous system through threshold registration algorithm. Results The three dimensional structures of the pulmonary venous system were reconstructed entirely. All reconstructed structures could be displayed individually or jointly, and could be rotated continuously on any plane. Conclusion The branches and the spacial characters of the pulmonary venous system can be clearly shown on the reconstructed 3D model. It will help anatomy teaching and provide morphological data for image diagnosis and lung operations. Related Articles | Metrics

技术方法

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AN COMPUTERIZED 3D RECONSTRUCTION METHOD BASED ON RAT’S CERVICAL CORD SERIAL SECTIONS 2008, 39 (1):  131-134.  doi: Abstract ( )   Objective With personal computer (PC) as the platform, to explore an computerized 3D reconstruction method based on rat’s cervical cord serial sections. Methods Normal cervical cord segment of SD rat was embeded in paraffin block, four locating marks were made at four corners around the specimen, the specimen was sectioned serially with Leica paraffin microtome, after each three sections, the images of specimen and outer locating marks were taken with CANON digital camera and the serial sections were obtained. The obtained images were ordered, cut, turned into gray scale images and background uneven corrected. The edges of white matter, grey matter and locating marks in each image were extracted, the locating marks were used for automated registration of serial sections with 3DDOCTOR software, the 3D surface reconstruction of cervical cord segment was observed, cut and measured freely in PC. Results The grey matter, white matter of rat cervical cord segment were reconstructed from serial transverse sections, the reconstructed segment was measured to obtain the surface area and volume data.Conclusion The paraffin embedding and “hard” locating marks techniques can be used to obtain the images of serial transverse sections of rat’s cerival cord segment, to make 3D reconstru Related Articles | Metrics

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DNA INTERFERENCE——THE REGULATION OF GENE EXPRESSION BY PROMOTERLESS HOMOLOGOUS DNA FRAGMENTS 2008, 39 (1):  135-136.  doi: Abstract ( )   Related Articles | Metrics