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2010, Volume 41 Issue 03
06 June 2010 Previous Issue    Next Issue
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Expression of glutamate receptors and GABASUB>A/SUB> receptors in the hippocampus of APPswe transgenic mice 2010, 41 (03):  333-338.  doi: 10.3969/j.issn.0529-1356.2010.03.001 Abstract ( )   Objective In order to understand the relationship between the receptors of excitatory and inhibitory neurotransmitter and Alzheimer disease(AD), the expression of glutamate receptors and GABASUB>Aα1-/SUB>SUB>6 /SUB>in the CA3 region of hippocampus of APPswe transgenic mice was investigated. Methods These mice from postnatal day 0 to postnatal day 360 were used in the study with the N-methyl-D-asprtate receptor 1(NMDAR1), GluR2/3 and GABASUB>Aα1-6/SUB> immunofluorescent stainings. In the meantime, Western blotting was carried out to detect the expressions of NMDAR1 and GABASUB>Aα1-6/SUB> in the hippocampus as well. Results The numbers of NMDAR1,GluR2/3 and GABASUB>Aα1-6 /SUB> positive neurons increased obviously after P7, in both wild type and transgenic mice, with the peaks at P30 However, in aged animals(P360), NMDAR1 and GluR2/3 lost in transgenic mice with statistically significant difference ( P< 005), comparing with wild type, but the alterations of GABASUB>Aα1-6/SUB> receptor was not obvious (EM>P/EM>>0.05) between wild type and transgenic mice. Conclusion The study showed that glutamate receptors in hippocampu Related Articles | Metrics

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Differentiation of bone marrow stromal cells into neurospheres induced by basic fibroblast growth factor combined with epidermal growth factor in rats 2010, 41 (03):  339-343.  doi: 10.3969/j.issn.0529-1356.2010.03.002 Abstract ( )   P>Objective To observe whether bone marrow stromal cells(MSCs) can be induced to form neurospheres by basic fibroblast growth factor(bFGF) combined with epidermal growth factor (EGF) in rats. Methods MSCs were cultured with whole bone morrow. Effect of bFGF and EGF, inducing MSCs to differentiated into neurospheres was studied by morphology, immunofluorescence, and Western blotting. Results MSCs can be effectively induced to form neurospheres by bFGF and EGF.The neurospheres can passage and differentiated into neural cells.Immunofluorescence and Western blotting showed these neurospheres expressed nestin protein. Cells differentiated from neurospheres expressed marker proteins of neurons, glial cells and oligodendrocytes. Conclusion The results indicate that MSCs can be effectively induced to form neurospheres by bFGF and EGF. The neurospheres may amplify and differentiate into neurons, glial cells and oligodendrocytes./P> Related Articles | Metrics

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Changes of microglia and astrocyte in substantia nigra of 6-OHDA model rats of Parkinson disease 2010, 41 (03):  344-348.  doi: 10.3969/j.issn.0529-1356.2010.03.003 Abstract ( )   Objective To investigate the changes of microglia and astrocyte in the substantia nigra of the rat models of Parkinson disease induced by 6-hydroxydopamine(6-OHDA). Methods Adult Sprague-Dawley rats were microinjected with 6-OHDA solution(8μl, 2mg/L) into the substantia nigra and medial forebrain bundle of the right side of the brain. After 2 weeks，there were 16 model rats, sixteen PD rats were randomly divided into two groups： 2-week and 8-week group, 8 in each group. Six adult normal rats were injected saline instead of 6-OHDA as the control group. Two weeks or 8 weeks later,the morphological changes of the dopaminergic neuron(tyrosine hydroxylase positive, TH-positive), the OX-42(specific marker for microglia)positive and GFAP(specific marker for astrocyte)positive in rat substantia nigra were examined by using immunohistochemical and immunofluorescent methods. Results There were only a few TH-positive cells in the 6-OHDA injected side of the model group. The differences between the ratios of the TH-positive cells of 6-OHDA injected side to the normal side in the two model groups and to the both sides in the control group were significant, respectively( P<0.01). There was no difference between the ratios of the two model groups( EM>P/EM>>0.05). The differences between the ratios of the number of the OX-42-positive cells and GFAP-positive cells between the 6-OHDA injected sides and the normal side in the two PD model groups are significant, respectively( EM>P/EM><0.01). The cell bodies of the OX-42-positive cells in 6-OHDA injected sides were ameboid in two PD model groups, and the processes of the GFAP-positive cells in 6-OHDA injected sides of the two model groups became shorter and deepen in color. There was no difference between the the ratios of those of the two model groups( EM>P/EM>>0.05). Conclusion The activation of microglia and astrocyte in the substantia nigra in Parkinson disease model by microinjection of 6-OHDA solution had been showed both in 2-week and 8-week model groups. There were no differences between the activation level of microglia and astrocyte in different time(2-week and 8-week mod Related Articles | Metrics

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Effect of heroin dependence on the pituitary adrenocorticotrophic hormone-positive cells and serum cortisol of rat 2010, 41 (03):  349-352.  doi: 10.3969/j.issn.0529-1356.2010.03.004 Abstract ( )   Objective To investigate the effect of heroin dependence on the expression of adrenocorticotrophic hormone （ACTH） in ACTH positive cells of pars distalis hypophyseos and the level of serum cortisol for exploring the possible mechanism of the changes. Methods Fifty-five adult male SD rats were randomly divided into a normal control group (NCG),a saline control group (SCG) and a heroin dependence group (HDG). Heroin-dependent rat models were set up by the way of subcutaneous injection of heroin and the hypophysis were excised on the 10th, 17th, 24th, 31st and 38th days after models were set up.Immunohistochemical SABC method, imagine analysis and radioimmunoassay(RIA) were used in the research. Results Compared with the NCG and SCG, the immune staining intensity of ACTH cells in HGD was weakened. The average gray value was significantly higher ( EM>P/EM><0.05 ).The result of RIA showed that the level of serum cortisol in HDG decreased as compared with the NCG( EM>P/EM>﹤0.05 ). Conclusion During heroin dependence, the expression of ACTH positive cells diminished, the level of serum cortisol reduced. The morphological and functional changes suggest that the heroin inhibits the function of the pituitary-a Related Articles | Metrics

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Neural protective action of astragalus injection in intracerebral hemorrhage rats 2010, 41 (03):  353-357.  doi: 10.3969/j.issn.0529-1356.2010.03.005 Abstract ( )   Objective To study the protective effect of astragalus injection on neurons that surround intracerebral hemorrhage（ICH）in rats. Method ICH model was established by injecting collagenase Ⅶ into the brain of rat. These rats in every group were graded according to the neurological function after analepsia. Immunohistochemical staining technique was used to demonstrate expressions of Bcl-2 and Bax protein. The ultrastructure of neurons that surround intracerebral hemorrhage in rats was observed by transmission electron microscope. Results The positive expression of Bcl-2 in treatment group was higher than that in model group（P<0.01)；However the positive expression of Bax in treatment group degraded prominently (P<0.01). The protein ratio of Bcl-2/Bax increased. Comparing with 0 hour,6 hours treatment group after surgery, the positive expression of Bcl-2 and Bax in 24 hours treatment group after surgery had prominently difference（EM>P/EM><0.05). The positive expression of Bcl-2 in 0 hour treatment group and 6 hours treatment group had no prominently difference（EM>P/EM>＞0.05). The ultrastructure of neuron in normal group was common, however, it was destroyed seriously in model group. The ultrastructure of neuron in astragalus treatment group improved obviously compared with model group. The ultrastructure of neuron in 0 hour,6 hours treatment group after ICH was better than in 24 hours treatment group. Quantitation analysis of nerve synapse displayed that the number of nerve synapse increased prominently in treatment group than in model group（EM>P/EM><0.01)；Meanwhile, the synaptic cleft became narrow conspicuously（EM>P/EM><0.01）；The compact area behind synapse thickened obviously（EM>P/EM><0.01）. The visual parameters of nerve synaptosome in 24 hours treatment group after surgery were different compared with 0 hour,6 hour Related Articles | Metrics

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Expression of microtubule-associated protein-2 and neurofilament-200 in rat brain after sleep deprivation 2010, 41 (03):  358-361.  doi: 10.3969/j.issn.0529-1356.2010.03.006 Abstract ( )   P>Objective To study the expression of microtubule-associated protein-2(MAP-2) and neurofilament-200(NF-200) in cortex, hippocampus and amygdaloid body of rat after sleep deprivation(SD). Methods SD model was made by modified multiple platform method（MMPM）, rats were divided into sleep deprivation group(SD),tank control group(TC), cage control group(CC). The sleep deprivation group included seven time-length groups (sleep deprivation 6 hours,12 hours,1 day,2 days,3 days,5 days and 7 days). Five rats were assigned to each group. The expression of MAP-2 and NF-200 were detected by immunohistochemical method. Results At the 5th day and 7th day, we found that the density of MAP-2 and NF-200 positive neuron decreased in cortex, CA1,CA2,CA3,dentate gyrus and amygdaloid body（P0.05，P0.01). Conclusio Related Articles | Metrics

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Effects of panax notoginseng saponins on proliferation and differentiation of cultured hippocampal neural stem cells EM>in vitro/EM> 2010, 41 (03):  362-366.  doi: 10.3969/j.issn.0529-1356.2010.03.007 Abstract ( )   Objective To research the effects of panax notoginseng saponins on proliferation and differentiation of rat hippocampal neural stem cells (NSCs) EM>in vitro/EM> . Methods NSCs of SD rat hippocampus were isolated and cultured in serum.free medium containing bFGF and EGF. The neurospheres were identified by antibodies of nestin, 5.bromodeoxyuridine (BrdU), class β.Ⅲ Tubulin(Tuj-1)and glial fibrillary acid protein (GFAP). The cells were divided into control group, PBS group and PNS treated group, which includes 0.1,0.2,0.4,0.8,1.0 g/L treated group. The clone information rate, proliferation curve and differentiation proportion of NSCs were observed. The cells were divided into control group and 0.4 g/L PNS treated group. Flow cytometry was used to study the effect of PNS on differentiation of hippocampal neural stem cells. Results The NSCs formed neurospheres and grew in floating. They were nestin-positive and BrdU-positive. GFAP-positive and Tuj-1-positive cells appeared after fetal bovine serum(FBS) were added into the medium. Compared with control, the low doses of panax notoginseng saponins (0.1,0.2,0.4 g/L) exhibited no effects on the NSCs proliferation, while high doses of panax notoginseng saponins (0.8 and 1.0 g/L) treatment markedly inhibited the proliferation of rat hippocampal NSCs and the difference had statistical significance ( P<0.05). Treatment of NSCs with 0.4 g/L PNS markedly increased the neurogenesis of NSCs in differential medium, and the difference had statistical significance( EM>P/EM><0.05). Conclusion Panax notoginseng saponins regulates the proliferation and differentiation of NSCs EM>in vitro/EM> , whereas only 200 mg/L panax notoginseng saponins can increase the neurogenesis of rat hippocampal NSCs. Related Articles | Metrics

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Protective effect of the decoction of angelicae sinensis for enriching blood on the cerebral cortex neurons of the focal cerebral ischemia reperfusion rats 2010, 41 (03):  367-373.  doi: changmanzhou@hotmail.com Abstract ( )   Objective To investigate the protective effect of the cerebral cortex neurons with the decoction of angelicae sinensis for enriching blood on the focal cerebral ischemia-reperfusion rats. Methods Forty-eight male SD rats were randomly divided into sham operation group, middle cerebral artery occlusion and reperfusion group,normal saline control group, decoction-treatment group. Before being operated, treatment-group rats were lavaged for 6 days, twice a day(taking medicine once in the morning and once in the evening ). Four hours after being operated，the rats were lavaged twice each 2.5ml. Rats will be sacrificed the following day.The defects of neurological behavior of the rats and the changes of cerebral infarct volume with TTC staining were also observed; 2% the Evans blue was injected into the femoral vein of rats to observe permeability changes of blood-brain barrier; The numbers and morphological change of neurons in the cerebral cortex were counted through Nissl staining; The p53 and Caspase-3 expression of the neurons in the cerebral cortex were detected with immunohistochemical method; Meanwhile, The apoptotic neurons in the cerebral cortex were observed through terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL) staining. Results When compared with the middle cerebral artery occlusion/reperfusion (MCAO) and control saline group, the infarct size in the decoction-treatment group was markedly smaller than that observed in TTC-staining (EM>P/EM><0.05).When brain slices were observed under a fluorescence microscope, less EB spillover was observed in the decoction-treatment group(EM>P/EM><0.05).The number of neurons in the cerebral cortex increased as observed by Nissl-staining(EM>P/EM><0.05). The number of positive immunohistochemistry staining of Caspase-3 and p53 was reduced (P<0.05). The number of apoptosis cell in the cerebral cortex was also reduced (P<0.05). Conclusion The decoction-treatment group could reduce death of neuron in the cerebral cortex. It protects neurons through the reduction of its target genes such as Caspase-3 and p53. Related Articles | Metrics

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Isolationculture and characteristics of human olfactory mucosa cells 2010, 41 (03):  374-378.  doi: 10.3969/j.issn.0529-1356.2010.03.009 Abstract ( )   Objective To establish a technical method to isolate and culture human olfactory mucosa cells EM>in vitro/EM>, and to study the immunological characteristics of olfactory ensheathing cells(OECs). Methods In this study, seven adult olfactory mucosae were obtained from nasal cavity surgery.Cells were acquired by dissection of olfactory mucosa and purified according to the different attachment time among the various cell types. The cells were observed for morphological characteristics under inverted microscope and identified using immunofluorescence staining with p75,S100 and glial fibrillary acidic protein(GFAP). The characteristics of olfactory ensheathing cells was observed under confocal microscope. Results S100 and p75 positive cells were more common in the olfactory mucosa cells under confocal microscope, GFAP positive cells were relatively rare. In general, S100 and p75 were co-expressed in the same cells, but not the GFAP and S100, p75 expressions. The p75 positive cells presented two completely different morphology. The percentage of S100 and p75 double labeled cells was (35.0±8.3)% under confocal microscope. Conclusion The olfactory ensheathing cells can be isolated from human olfactory mucosa,they are accessible sources for autologous grafting to repair nervous sys Related Articles | Metrics

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Effect of silencing Huntingtin-associated protein 1 on the apoptosis of mouse islet beta cells line NIT cells 2010, 41 (03):  379-382.  doi: 10.3969/j.issn.0529-1356.2010.03.010 Abstract ( )   Objective To investigate the effect of silencing Huntingtinassociated protein 1（HAP1）on apoptosis of mouse pancractic islet beta cells line(NIT cells). Methods 1.The NIT cells were transfected with chemically synthesized siRNA targeting HAP1 for comprehensive evaluation of its silence efficacy. 2.The expression of the silencing HAP1 was detected by Annexin V and PI, and TdT-mediated dUTP-biotin nick end-labeling (TUNEL) to evaluate its efficacy on the apoptosis of NIT cells. 3. The expression of the silencing HAP1 was detected by Annexin V and PI to evaluate its efficacy on the streptozotocin (STZ)-induced apoptosis of NIT cells. Results The expression of silencing HAP1 in NIT cells was inhibited significantly by chemically synthesized siRNA targeting HAP1; The number of the apoptosis of NIT cells based on estimates of the difference between experimental and control group, it was markedly increased in the group transfected with chemically synthesized siRNA targeting HAP1 as compared with the control group (P<0.01); STZ-induced apoptosis of NIT cells was significantly increased (P<0.01), and the expression of silencing HAP1 was sufficient to promote STZ-induced apoptosis of NIT cells (P<0.01). Conclusion The expression of silencing HAP1 increases the apoptosis of mice pancreatic islet beta cells line NIT, and it enhances the capability of apoptosis-inducer, STZ and to quicken the apoptosis of NIT cells in s Related Articles | Metrics

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Effects of prolonged starvation on aminotransferases, phosphatases, proteinases and proliferating cell nuclear antigens in planarians 2010, 41 (03):  383-386.  doi: 10.3969/j.issn.0529-1356.2010.03.011 Abstract ( )   Objective To investigate the effects of prolonged starvation on aminotransferases, phosphatases, proteinaes and proliferating cell nuclear antigens (PCNA) in planarians. Methods Automated biochemistry analyzer was employed to determine the enzymes activities of aminotransferases, non-denatured electrophoresis (G-PAGE) was used to analyze enzymes activities of phosphatases and proteinaes, the semi-quantitative RT-PCR was employed to investigate the expressed changes of PCNA mRNA. Results The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) elevated apparently during starvation, 10-fold higher than that of the control level, but reduced to the normal level after re-fed; The activities of alkaline phosphatase (ALP) decreased during starvation, whereas the activities of acid phosphatase (ACP) increased apparently; In response to starvation, the activities of 140kD and 40kD proteinases increased drastically; However, no changes of the expression level of PCNA mRNA during starvation were observed, which indicated that starvation had little influences on this gene. Conclusion Prolonged starvation has greater effects on the physiology and biochemical metabolism of planarians, and the changes of enzyme activities reflecte the changes of energy metabolism of planarians in response to prolonged starvation. This findings provide a basis for further understanding the physiological mechanism that planarians are in adaptation to starvation. Related Articles | Metrics

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Eexogenous spermidine and spermine induce the expression of antizyme in early regenerating rat liver 2010, 41 (03):  387-390.  doi: 10.3969/j.issn.0529-1356.2010.03.012 Abstract ( )   Objective To study the roles of spermidine and spermine in affecting antizyme expression in early regenerating rat liver. Methods Adult male SD rats were surgically rendered partial hepatectomy via a modification of a procedure reported by Higgins and colleagues (1931), then they were treated with different dosages of exogenous polyamines (dissolved in 0.9% NaCl) s.c. Antizyme (AZ) gene transcription and protein expression in regenerating liver were evaluated by using the RT-PCR and Western blotting assay respectively. Results In intact liver (0 hour after partial hepatectomy) of the control group, the levels of AZ mRNA transcription and protein expression were very low. In regenerating liver induced by partial hepatectomy, AZ mRNA content and protein expression increased quickly, peaked at 3 hours, followed by significant decreases at 5 hours, afterwards, it reached another peak at 7 hours, and then decreased gradually. In rats administered with different doses of spermidine and spermine, the change tendency of AZ mRNA level and protein expression in regenerating liver paralleled with those of control group. But the levels in low-dose-treated-group were greatly lower than that in high-dose-treated-group at each time point throughout the whole experiment. The actions of spermine were more obvious and longer. Conclusion Exogenous polyamines can induce antizyme express dose-dependently in early regenerating of rat liver, and the effects of spermine were more powerful than that of spermidine. Related Articles | Metrics

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Expression level and amplification condition of HER2 and TOP2A as well as their relationship in breast cancer 2010, 41 (03):  391-394.  doi: 10.3969/j.issn.0529-1356.2010.03.013 Abstract ( )   Objective To investigate the expression level and amplification condition of HER2 and TOP2A as well as their relationship in breast cancer. Methods Immunohistochemistry and fluorescent in situ hybridization (FISH) were taken to detect the the expression level of HER2/topoⅡα, amplification status of HER2 gene/TOP2A gene respectively. Statistical analysis was performed to explore the correlation between them. Results The breast cancer tissues of 58 patients were tested. The expression level of HER2 was investigated in 11 cases with negative expression（11/58,19.0%）,19 cases with weakly positive expression（19/58,32.8%），13 cases with positive expression（13/ 58,22.4%）and 15 cases with strongly positive expression（15/58,25.8%）. The expression level of topoⅡα was investigated in 28 cases with negative expression（28/58,48.3%），22 cases with positive expression（22/58,37.9%），8 cases with strongly expression（8/58,13.8%）. Amplification of HER2 and TOP2A gene was detected in 19 patients（19/58,32.8%）, 11 patients（11/58,19.0%）respectively. Non-amplification of HER2 gene and TOP2A was detected in 39 patients （39/58,67.2%）, 47 patients（47/58,81.0%）respcetively. Spearman rank correlation analysis revealed that the expression level of HER2 protein was significantly related with the amplification status of HER2 gene (EM>rSUB>s/SUB>/EM>=0.80,EM>P/EM><0.05）and the expressio Related Articles | Metrics

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Epigallocatechin-3-gallate inhibits the expression of matrix metalloproteinase-1 in mouse cutaneous wound healing 2010, 41 (03):  395-399.  doi: 10.3969/j.issn.0529-1356.2010.03.014 Abstract ( )   Objective To explore the effects of epigallocatechin-3-gallate(EGCG) (50 mg/L) on the expression of MMP-1 mRNA and the levels of MMP-1 during mouse cutaneous wound healing. Methods Fifty-seven mice were divided into control, 50 mg/L EGCG and vehicle groups. The wound skins were prepared at 6, 12, 24, 72, 120 and 168 hours after injury, respectively. The expression of MMP-1 mRNA and levels of MMP-1 in wound healing skins were determined with reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). Results It was found that 50 mg/L EGCG inhibits the expression of MMP-1 mRNA and decreases the levels of MMP-1 of the wound tissue at the late stage of wound healing (72 hours after skin injury). Conclusion 50 mg/L EGCG could inhibit the expression of MMP-1 mRNA and the MMP-1 production during cutaneous wound healing, a Related Articles | Metrics

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P>Effect of manganese on Caspase-3 mRNA regulation in spermatogenic cell and the expression of vimentin on Sertoli cell in rats/P> 2010, 41 (03):  400-404.  doi: 10.3969/j.issn.0529-1356.2010.03.015 Abstract ( )   Objective To determine the effect of manganese on Caspase-3 mRNA regulation in spermatogenic cell and the expression of vimentin on Sertoli cell in rat. Methods Forty-eight male rats were randomly divided into one control group and two manganese chloride(MnCl-2)(15mg/kg,30 mg/kg ) groups. After exposed to manganese for 4 and 6 weeks respectively, the apoptosis of spermatogenic cell was examined by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL) technique. The expression of Caspase-3 mRNA in spermatogenic cells was investigated byEM>in situ/EM> hybridization,and the expression of vimentin on Sertoli cell of rats was detected by immunohistochemitry(SABC). Results 1. Compared with control groups, AI was increased(EM>P/EM><0.05，EM>P/EM><0.01)， the Caspase-3 mRNA-positive-cell rate was increased significantly ( P<0.01)， the vimentin-positive-cell rate was decreased significantly(P<0.01) in every manganese exposure groups. 2. Among manganese exposure groups of same dose, AI and the Caspase-3 mRNA-positive-cell rate were increased more significantly, but the vimentin-positive-cell rate was decreased more significantly in 6weeks manganese exposure groups than that in 4weeks manganese exposure groups (EM>P/EM><0.01). 3. Among manganese exposure groups of same time, AI and Caspase-3mRNA-positive-cell rate in 30mg/kg manganese exposure groups were increased more significantly than that in 15mg/kg manganese exposure groups ( EM>P/EM><0.01), but the vimentin-positive-cell rate was decreased more significantly in 30 mg/kg manganese exposure groups than that in 15 mg/kg manganese exposure groups ( EM>P/EM><0.01). 4. There existed a positive correlation between AI and the Caspase-3 mRNA-positive-cell rate (EM> r/EM> =0.842, EM>P/EM><0.01)， a negative correlation between AI and vimentin-positive-cell rate (EM>r/EM>=-0.859, EM>P/EM><0.05)，and a negative correlation between the Caspase-3 mRNA-positive-cell rate and the vimentin-positive-cell rate (EM>r/EM>=-0.975, EM>P/EM><0.01). Conclusion After exposed to manganese the increased expression of Caspase-3 mRNA in spermatogenic cells led to increased apoptosis of spermatogenic cell and decreased express Related Articles | Metrics

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Determination and analysis of eight seroenzymes biochemical indexes of panthera tigris amureusis 2010, 41 (03):  405-408.  doi: 10.3969/j.issn.0529-1356.2010.03.016 Abstract ( )   Objective The eight enzymes’ levels in serum from Panthera tigris amureusis （ P. t. amureusis ） were determined. The data wouldl be accumulated for the growth, reproduction and disease diagnosis and treatment of young P. t. amureusis , as well as types of tiger subspecies genetic and evolutional relationship studies. Methods Eight enzymes’ biological indexes of 13 individuals had been measured and analyzed in P. t. amureusis by HITCH 7200-20 automated chemistry analyzer, respectively between male and female young P. t. amureusis , and between young P. t. amureusis and adult. Results Aspartate aminotransferase(AST) was (29.31±5.88)IU/L, lactate dehydrogenase enzyme(LDH) was (267.08±76.40)IU/L,α-hydroxybutyrate dehydrogenase(α-HBDH) was (149.38±54.07)IU/L, gamma-glutamyl transpeptidase(GGT) was (2.92±1.94)IU/L, creatine kinase(CK) was (284.77±132.02)IU/L, creatine kinase isoenzyme(CKMB) was (390.62±145.70)IU/L and actate dehydrogenase isoenzyme(LDI) was (11.11±7.08)IU/L.Conclusion There are no differences in Related Articles | Metrics

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Expressions of 14-3-3ζ and E-cadherin proteins in stage TSUB>1/SUB> non-small cell lung cancer and the clinical significance 2010, 41 (03):  409-413.  doi: 10.3969/j.issn.0529-1356.2010.03.017 Abstract ( )   Objective To observe the expressions of 14-3-3ζ and E-cadherin in stage TSUB>1/SUB> non-small cell lung cancer(NSCLC) in order to investigate the relatioship between expressions of these two kinds of proteins and development and invasion of lung cancer. Methods Specimens of NSCLC and adjacent normal lung tissues were collected from 110 patients. Expressions of 14-3-3ζ and E-cadherin were detected by immunohistochemistry and Western blotting. Results Up-regulation of 14-3-3ζ was observed in stage in NSCLC tissue. The increased 14-3-3ζ protein levels were associated with differentiation degree and lymph node metastasis and it was no relationship with age,gender and histological type. Decreased expression of E-cadherin was observed in NSCLC tissue. The expression was related to differentiation degree and lymph node metastasis and it was no relationship with age,gender and histological type. Conclusion The increased expression of 14-3-3ζ contributes to NSCLC development and progression. Related Articles | Metrics

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Effect of trichosanthin on the first meiosis of mouse oocytes 2010, 41 (03):  414-418.  doi: 10.3969/j.issn.0529-1356.2010.03.018 Abstract ( )   Objective To investigate the effect of trichosanthin（TCS） on the first meiosis of mouse oocytes. Methods Totally 480 immature (GV-stage) oocytes were collected from the female mice (7-9 weeks old) at the 48th hour after they were received superovulation with 10IU of pregnant mare serum gonadotropin （PMSG）. They were cultured in the medium containing different concentrations of TCS. We observed the germinal vesicle breakdown (GVBD) percentage under the inverted microscope (anatomical lens). We detected the spindle formation and the chromosomes movement by immunofluorescence staining and then counted the number of the oocytes progressed to metaphaseⅠ(MⅠ) with laser scanning confocal microscope (LSCM). Oocytes were collected and observed to count the percentage of the first polar body (PB1) extrusion 16 hours after culture, served as the mark of the completion of the first meiosis. Results It was no significant deviation that the percentage of GVBD incidence and PB1 extrusion in the presence of TCS (30mg/L) compared with the control group; The lower concentration of TCS (30mg/L) interfered with the spindle assembly and the chromosomes alignment, which caused a significant decrease in the number of the oocytes progressed to MⅠand then extruded PB1; The higher concentrations of TCS (120-240mg/L) resulted in a significant decrease in the percentage of oocytes that underwent GVBD. Conclusion TCS interferes with the resumption of the meiosis, the spindle assembly and chromosomes alignment. And t Related Articles | Metrics

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Characteristic comparison of human embryonic stem cells in serum-containing and serum-free medium 2010, 41 (03):  419-424.  doi: 10.3969/j.issn.0529-1356.2010.03.019 Abstract ( )   Objective To evaluate the biological characteristic of human embryonic stem(hES) cells in serum-containing and serum-free culture condition. Methods Human embryonic stem cell line BG02 was cultured on mitomycin C treated mouse embryonic fibroblasts(MEF) feeder layers in medium supplied with serum or knockout serum replacement for 25-30 passages to compare their morphology. The proliferation of hES cells clones was detected by BrdU incorporation assay. The double time of hES cells was evaluated by cell counting. The pluripotency markers of hES cells were examined by immunofluorescence staining. The proportions of Oct-4 and Nanog positive cells were assessed by flow cytometry. Expressions of fibroblast growth factor(FGFs) family genes were determined by RT-PCR. Results Although BG02 cells in the serum and serum-free culture medium shared similar morphology, hES clones in serum-free culture condition showed more typical morphology and more Oct-4, Nanog positive cells than that in serum-containing condition. The attachment rate of hES cells clumps cultured in serum-free condition was obviously higher than that of serum condition ( EM>P /EM><0.05). The growth of BG02 cells in serum-containing medium was more fast (EM> P/EM><0.05). The doubling time of hES cells in the serum and serum-free condition was (45.9±5.7)hours and (33.8±4.3) hours, respectively. Additionally, BG02 cells in serum-free culture condition expressed high level of FGF2,FGFR2, and FGFR4. Conclusion BG02 cells cultured in serum or serum-free culture condition represent some variations in the growth and characteristics of hES cells. These variations might be attributed to the activation of members of FGFs family in BG02 cells. Related Articles | Metrics

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Degradation pattern and element distribution of WE43-magnesium alloy implanted in rats 2010, 41 (03):  425-429.  doi: 10.3969/j.issn.0529-1356.2010.03.020 Abstract ( )   Objective To investigate the feasibility of WE43 magnesium alloy applied as balanced degradable fixed materials in the bones of human body. Methods Both experimental WE43 and control Mg-Mn alloy rods were implanted into the same femora of 18 SD rats, which were sacrificed at different time postoperatively for hepatic and renal pathological analysis, implant/bone interface observation with stereoscope and scanning electron microscope(SEM), degradation rate counting and elements distribution scanning. Results WE43 rods showed better corrosion resistance and osteoinductive properties than Mg-Mn , although both of them degraded simultaneously. Likewise, both materials scarcely harmed liver and kidney of rats. A few earth elements of WE43 rod remained in the new bone and could promote bone development, while Mg and Mn elemnts in whatever alloy were not existed in the peripheral bone tissue. Conclusion WE43 alloy degrades slower than Mg-Mn alloy, which enables it to adapt to bone forming and rebuilt balance relationship between them. Therefore, WE43 is a potential biodegradable bone implant with good histocompatibility. Related Articles | Metrics

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Attachment and proliferation of induced osteoblasts from bone marrow stromal stem cellsEM> in vitro/EM> on degradable magnesium alloy 2010, 41 (03):  430-434.  doi: 10.3969/j.issn.0529-1356.2010.03.021 Abstract ( )   Objective To investigate the attachment and proliferation of osteoblasts from bone marrow stromal stem cells (BMSCs) on the surface of Mg-Mn-Zn alloy. Methods Rabbit BMSCs were isolated by density gradient centrifugation method and amplified in the flasks, using the osteogenic inducing conditions (OGC) as the culture media. The osteogenic potential of BMSCs was investigated by means of Alizarin red staining, alkaline phosphatase (ALP) staining, collogenⅠimmunohistochemisty technology , scanning electron microscope(SEM) and transmission electron microscope(TEM). Besides, induced osteoblasts were seeded on the surface of Mg-Mn-Zn alloy and pure Ti by 1×10SUP>5/SUP>/ml. Scanning electron microscopy and laser scanning confocal microscopy were carried out to observe cell morphology and growth at 1, 3 and 5 days under co-culture condition. Results Rabbit BMSCs growed in a great number and in the form of cell spheroids and the cells outgrew in all directions firstly. These cells were then induced into osteoblasts which could adhere to the surface of Mg-Mn-Zn and connected with each other to form lamellar substances. Conclusion Osteoblasts from BMSCs have good adhesion and growth capability on the surface of Mg-Mn-Zn indicating that magnesium alloy posses good bone induction function. Related Articles | Metrics

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Effect of low-dose gossypol acetic acid combined with steroid hormones on the expressions of Cdc25A,cyclin B1 and androgen receptor in adult rat testis 2010, 41 (03):  435-439.  doi: 10.3969/j.issn.0529-1356.2010.03.023 Abstract ( )   Objective To investigate the expressions of Cdc25A,cyclin B1 and androgen receptor (AR) by administering with combination regimen of low-dose gossypol acetic acid(GA) with steroid hormones ［desogestrel/ethinylestradiol/testosterone undecanote(DSG/E/TU)］ in adult rat testis． Methods Adult male rats were randomly divided into four groups．Group GH:rats were fed orally with GA［12.5mg/(kgI>&#/I>8226;day)］ and DSG［125μg/(kgI>&#/I>8226;day)］/E［25μg/(kgI>&#/I>8226;day)］/TU［100mg/(kgI>&#/I>8226;day)］;Group G:rats were administered with a single dose of GA［12.5mg/(kgI>&#/I>8226;day)］;Group H:DSG［125μg/(kgI>&#/I>8226;day)］/E［25μg/(kgI>&#/I>8226;day)］/TU［100mg/(kgI>&#/I>8226;day)］were administered to rats;Group C:rats only received vehicle(1% methyl cellulose)．The testes were excised at the 6th，8th and 10th week, respectively.The expressions of Cdc25A,cyclin B1 and AR are detected by immunohistochemical staining and Western blotting． Results Compared with the control group, the intensities of Cdc25A immunohistochemical positive staining were decreased in Leydig cells and acrosomes at the 6th，8th and 10th week(EM>P/EM><0.05), and increased in spermatocytes in G，H and GH group at the 6th and 8th week (EM>P/EM><0.05). The intensities of cyclin B1 immunohistochemical positive staining were decreased in Leydig cells in H and GH group at the 6th，8th and 10th week(EM>P/EM><0.05). AR positive staining appeared in the nuclei of Leydig cells in H and GH group instead of the cytoplasm. Western blotting showed that the expre Related Articles | Metrics

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Effect of Heshouwuyin on Rb/p53 signal transduction pathway in aging rat testis tissue cells 2010, 41 (03):  440-445.  doi: 10.3969/j.issn.0529-1356.2010.03.022 Abstract ( )   Objective To investigate the mechanism of Heshouwuyin in anti-aging in testis of the aging model rats by detecting expression of Rb,p16, p53, p19 and p21 in aging rats testis. Methods Totally 84 of 8 month male SD rats were selected and randomly divided into normal group, model group, Heshouwuyin high，middle，low group, Heshouwuwan group and spontaneous recovery group with 12 rats in each group The sub-acute aging rats were made by injecting D-gal into abdominal cavity continually. SP immunohistochemistry, Western blotting and RT-PCR were used to observe the differential expression of phosphorylated Rb protein, p16, p53, p19 and p21 in testis tissue. Results The expression of testis tissue Rb, p53, p16, p19 and p21 in model group rat obviously increased, the expression of pRb obviously decreased (P<0.05); Heshouwuyin and Heshouwuwan group raised the expression of pRb, decreased the expression of Rb, p53, p16, p19, p21(EM>P/EM><0.05), and the regulating effect of Heshouwuyin high group was more effective than rest groups(EM>P/EM><0.05). Conclusions Heshouwuyin regulates the expression of Rb/p53 signal transduction pathway related gene and protein in testis tissue to Related Articles | Metrics

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Stereological quantification of capillaries in rat white matter and cortex 2010, 41 (03):  446-451.  doi: 10.3969/j.issn.0529-1356.2010.03.024 Abstract ( )   Objective To study the total length, total volume, total surface area and average diameter of the capillaries in rat white matter and cortex. Methods In the present study, 5 adult LongEvans rats (7 months) were used. The length density, volume density, surface area density and total length, total volume, total surface area, average diameter of the capillaries in white matter and cortex were estimated with the stereological techniques and immunohistochemistry technique. Results The tissue processing induced volume shrinkage of rat white matter (±s) was 0.51 ± 0.09. The total length, total volume, total surface area and average diameter of the capillaries in the white matter was (30.9 ± 6.2) m，(0.8±0.09) mmSUP>3/SUP>，(5.3±0.74)cmSUP>2/SUP>，(5.5±1.1)μm. The tissue processing induced volume shrinkage of rat cortex was 0.57±0.08. The total length, total volume, total surface area and average diameter of the capillaries in the cortex was (364.8±68.0)m，(7.0±1.6)mmSUP>3/SUP>，(51.6±11. Related Articles | Metrics

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Correlation between MR-DWI and the expression of aquaporin-4 after acute cerebral ischemia and reperfusion in rabbits 2010, 41 (03):  452-457.  doi: 10.3969/j.issn.0529-1356.2010.03.025 Abstract ( )   Objective To disclose the relationship between aquaporin-4（AQP-4）and apparent diffuse coefficient （ADC） by analyzing the magnetic resonance diffusion weighted imaging（MRI-DWI） of acute cerebral ischemia and reperfusion in rabbits. Methods Fifty- eight New Zealand white rabbits were divided into permanent ischemia group(A group, EM>n/EM>=30)which were subdivided into 1 hour, 3 hours, 6 hours, 12 hours, 24 hours, 48 hours groups, and reperfusion group after 1 hour cerebral ischemia (B group, n=28)which were subdivided into 0 hour, 2 hours, 5 hours, 11 hours, 23 hours, 47 hours groups. All rabbit brains were cut out to make histologic section immediately after each time point MRI examination. AQP-4 detection was carried out in bilateral cortical and corpus striatum region respectively under microscope. ADC and rADC value and positive cells of AQP-4 were calculated and recorded. Results The hyperintensity area in MRI-DWI tended to enlarge gradually in permanent ischemia groups. The relative apparent diffusion coefficient （rADC） value decreased at first and then increased gradually. In reperfusion groups, the rADC value increased at the 2nd hour point with shrinkage of hyperintensity area on DWI.After 5 hours of reperfusion, the rADC value increased again with enlargement of DWI hyperintensity area. AQP-4 positive cells decreased within 6 hours, and then went up in permanent ischemia groups. There was a close relationship between rADC value and AQP-4 positive cells(EM>r/EM>=0.943, EM>P/EM>=0.005). The same things happened in reperfusion groups within 5 hours, but the scope of chan Related Articles | Metrics

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Three-dimensional reconstruction of hippocampal formation in normal adult mice 2010, 41 (03):  458-461.  doi: 10.3969/j.issn.0529-1356.2010.03.026 Abstract ( )   Objective To make three-dimensional (3D) reconstruction of mouse brain and inner hippcampal formation (HF) with 3D software, and to observe and measure the reconstructed structures. Methods The Nissl stained mouse brain serial cryosections were obtained and pre-processed to construct the image dataset for 3D reconstruction. Using PhotoshopCS3 software, the different anatomic structures of HF in mouse coronary brain sections were fulfilled with different colors according to anatomy knowledge. The 3D-DOCTOR 4.0 software was used to make registration, segmentation and 3D reconstruction of mouse brain and HF, and to observe the reconstructed structures in personal computer (PC). Results The serial Nissl stained cryosections could be used to make 3D reconstruction of mouse HF, and the subiculum, CA1, CA2, CA3, CA4, dentate gyrus (DG), etc, could be observed. Conclusion The serial Nissl stained mouse cryosections could be used to make 3D reconstruction of mouse brain and HF. Related Articles | Metrics

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Three-dimensional reconstruction and morphometry method of rat kidney based on serial sections 2010, 41 (03):  462-468.  doi: 10.3969/j.issn.0529-1356.2010.03.027 Abstract ( )   Objective To explore an easy to perform three-dimensional reconstruction method based on serial section images of large tissue by combination of several new techniques. Methods After the processes of HE stain, digitalization, assembling and alignment, the digitalized image database of long serial sections from a normal rat renal paraffin-embedded tissue was made, in which the vascular system and pelvis were segmented and reconstructed, the morphology figures of constructed result were also measured. Results The image database composed of clear whole images and correct registration reappeared structural pattern of the tissue, and the results displayed the complicated positional relationship of renal structures and could be observed freely, some three dimensional figures including percentage components of kidney, length and angel of arteries were obtained. Conclusion We reported a three-dimensional reconstruction method from serial sections with accepting results. It can also be used in other cases needing stereoscopic information or intuitive observation. Related Articles | Metrics

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Expression of T-type calcium channels during mouse embryonic heart development 2010, 41 (03):  469-472.  doi: 10.3969/j.issn.0529-1356.2010.03.028 Abstract ( )   Objective To investigate the temporal and spatial expression pattern of the pore-forming α1 subunits of T-type calcium channels during mouse embryonic heart development. Methods Histology and immunohistochemistry of mouse embryonic hearts from embryonic 9.5 days （E9.5） to E18.5 in paraffinembedded were used to analyze the expression of α1G and α1H, two subtypes poreforming α1 subunits of T-type calcium channels. Results α1H proteins were detectable from earlier embryonic days (E10.5) to late embryonic day (E18.5) while α1G proteins from E11.5 to E18.5. Immunohistochemical localization showed that the expression of α1H or α1G throughout myocardium and bundle branches with exception of the endothelium and mesenchyme cells among the trabecular and endocardial cushion re Related Articles | Metrics

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Effect of RGD stimulating integrin of cardiomyocyteson the uptake of macromolecules 2010, 41 (03):  473-476.  doi: 10.3969/j.issn.0529-1356.2010.03.029 Abstract ( )   Objective To test whether stimulation of integrins on neonatal rat cardiomyocytes (NRCMs) would favor uptake of macromolecules. Methods NRCMs were treated with Gly-Arg-Gly-Asp-Ser (RGD, 100-300 mg/L) to stimulate integrins, or phosphate-buffered saline, in the presence of ovalbumin-texas red (OTR, 45 kD) or dextran-texas red (DTR, 70 kD). After 8 hours, 16 hours, 24 hours, uptake of red label as well as Dysferlin immunoreactivity were quantified by fluorescence microscopy. Results Uptake of OTR and DTR by NRCMs was intensified by RGD ( for trend EM>P/EM><0.001 and EM>P/EM><0.05, respectively) and was correlated with duration of incubation (EM>P/EM><0.001 for both). RGDinduced uptake of OTR by NRCMs was correlated with OTR concentration (EM>P/EM><0.001). The Dysferlin-positive areas in RGD-treated NRCMs were about 3-fold larger than in NRCMs incubated without RGD (EM>P/EM><0.001). However, in non-beating NRCMs RGD did not induce larger dysferlin-positive areas, nor did RGD stimulate uptake of OTR. Conclusion In NRCMs stimulation of integrins caused uptake of labeled macromolecules. Dysferlin may be involved in membrane repair following RGD-induced membrane wounding. RGD-induced membrane wounding is closely associated with contractile behavior of the NRCMs. Related Articles | Metrics

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Digit and digit ratio of Liaoning Han nationality 2010, 41 (03):  477-479.  doi: 10.3969/j.issn.0529-1356.2010.03.030 Abstract ( )   Objective To study Liaoning Han nationality’s characteristics of digit ratio. Methods With informed consent, we randomly chose 728 (270 males and 458 females) healthy Liaoning Han people between 20 and 22 years old, excluding patients of endocrine and metabolic disease and the people whose fingers were malformed and injured. Their mothers had never taken any hormones during duration of pregnancy. We used direct measurement to measure digit length, SPSS 14.0 package to statistical analysis. Results Liaoning Han people’s digit length present 3D＞4D＞2D＞5D，female’s 2D＞male’s(EM>P/EM><0.001）、male’s 4D>female’s（EM>P/EM><0.001）;Digit ratio of Liaoning Han people possessed the tendency of 3D:5D＞4D:5D＞2D:5D＞3D:4D＞2D:4D＞2D:3D; Digit ratio of Liaoning Han nationality had sexual and bilateral differences, especially in 2D:4D; Digit ratio of Liaoning Han people was exceed the digit ratio of Related Articles | Metrics

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A noval double-compound staining method showing Nissl bodies and myelin sheath in neuron 2010, 41 (03):  480-482.  doi: 10.3969/j.issn.0529-1356.2010.03.031 Abstract ( )   Objective To develope a method for staining Nissl bodies and myelin sheath in spinal cord tissues and the spinal ganglion from 8 New Zealand white rabbits. Methods The double-compound staining dyes composed of brilliant cresyl violet, toluidine blue,thionin,orange G and phosphotungstic acid (BTT-OP method) were investigated. Results The Nissl body,axons， nerve membrane and connective tissue were stained blue, the nerve cell nuclei were stained yellow, the myelin sheath and the background showed slight yellow. Conclusion Double-compound staining method （BTT-OP method ）possesses more vivid color and more striking contrast than the original method, it can demonstrate clearly the structures of neuron under the light Related Articles | Metrics