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2009, Volume 40 Issue 5
06 October 2009 Previous Issue    Next Issue
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EM>In vitro/EM> study of Akt2 function on U87 malignant glioma cells migration and invasion 2009, 40 (5):  691-695.  doi: 10.3969/j.issn.0529-1356.2009.05.001 Abstract ( )   Objective To detect the effect of Akt2 on U87 glioblastoma cells migration and invasion by Akt2 small RNA interference technology EM>in vitro/EM>.Methods Akt2 expression was knocked down in U87 glioblastoma cells by Akt2 siRNA, stable clones were used to perform wound healing assay, F-actin polymerization assay, adhesion assay and matrigel invasion assay. Western blotting was used to check the expression level of phosphorylated cofilin and LIMK. Results Result of wound healing assay suggested that motility of U87 cells was significantly decreased with deficiency of Akt2; U87 cells with Akt2 deficiency was not capable of assembling actin effectively compared to control cells by F-actin polymerization assay. Further, phosphorylated cofilin and LIMK expression which are important for cellular motility were all decreased with Akt2 deficiency. Besides, adhesion and invasion ability of U87 cells were significantly impaired after Akt2 was knocked down. Conclusion Akt2 is important for U87 cells migration and invasion. Akt2 deficiency will lead to impairment of U87 glioblastoma cells motility, adhesive and invasive ability EM>in vitro/EM>. Related Articles | Metrics

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Polypeptides of EM>Achyranthes bidentata/EM> Blume promotes the neurite growth of rat hippocampal neurons 2009, 40 (5):  696-701.  doi: 10.3969/j.issn.0529-1356.2009.05.002 Abstract ( )   Objective To determine the effects of polypeptides of Achyranthes bidentata Blume (ABPP) on neurite growth in cultured hippocampal neurons of rats. Methods The cell activity of rat hippocampal, neurons with ABPP was tested with MTT. After 24 hours of incubation with different concentrations (0.1 mg/L, 0. 5 mg/L, 1.0 mg/L)of ABPP, the hippocampal neurons were photographed by TCS SP2 confocal microscope with fluorescent immunocytochemistry, and the neurite length was analyzed using the Image-Pro Express software. Real-time fluorescence quantitative RT-PCR was performed to examine the mRNA levels of GAP-43 and NF-H after 6 hour with different concentrations of ABPP. Western blotting was performed to further examine the protein levels of GAP-43 and NF-H in cultured hippocampal neurons after 24 hours incubation with different concentrations of ABPP. Results ABPP promoted the neurite growth of cultured hippocampal neurons. Real-time fluorescence quantitative RT-PCR showed that the mRNA level of GAP-43 and NF-H in cultured hippocampal neurons was up-regulated by ABPP in a dose dependent manner. Meanwhile, fluorescent immunocytochemistry and Western blotting showed that the protein level of GAP-43 and NF-H were up-regulated by ABPP. Conclusion ABPP could promote the neurite growth and modulate the expression of GAP-43 and NF-H in cultured hippocampal neurons, which maybe have something to do with ERK pathway. Related Articles | Metrics

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A light and electron microscopic study on changes of dopamine receptors in the rat neostriatum after unilateral lesion of the nigrostriatal dopaminergic pathway 2009, 40 (5):  702-708.  doi: 10.3969/j.issn.0529-1356.2009.05.003 Abstract ( )   Objective There have been robust of reports about quantity changes of dopamine receptors but there has no report describing the distribution changes of dopamine receptors at ultrastructural level in Parkinson disease until now. As whether those receptors are functional and the magnitude of their function is closely related to their distribution, it is necessary to study their distribution changes at ultrastructural revel under the disease situation. Methods Changes of striatal dopamine D1 receptors (D1R) and D2 dopamine receptors (D2R) in rats 4 weeks and 16 weeks after 6-hydroxydopamine(6-OHDA) unilateral lesion nigro-striatal dopaminergic pathway were studied by immunohistochemical and pre-imbedding immunogold electron microscopic methods. Results Four weeks after lesion, striatal D1R ipsilateral to lesion side decreased as observed under light microscope. The decrease happened at spines but not dendrites or cell bodies under electron microscope. 16 weeks after lesion, although the degree of striatal D1R ipsilateral to lesion side returned to the degree contralateral to the lesion side at light microscopic level, the amount of D1R decreased at spines and dendrite but increased significantly at cell bodies at electron microscopic level. Both 4 and 16 weeks after lesion, striatal D2R increased ipsilateral to lesion side under light microscope, the increased D2R particles were found at spines and dendrites but not cell bodies. Moreover, more gold particles of the two receptors were found at the extrasynaptic site both 4 weeks and 16 weeks after lesion. Conclusion These changes might play important roles in the onset and development of Parkinson’s disease and mechanism of side effect of levodopa therapy. Related Articles | Metrics

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Effects of short-term enriched environment on the white matter and the myelinated fibers in the white matter of aged male rats 2009, 40 (5):  709-714.  doi: 10.3969/j.issn.0529-1356.2009.05.004 Abstract ( )   Objective To investigate the effects of short-term enriched environment on the white matter and the myelinated fibers in the white matter of aged male rats. Methods Twenty 24-month old male SD rats were reared in enriched environment (EE) or standard environment (SE) for 4 months, respectively. Four rats were randomly selected from each group. The white matter and the myelinated fibers in the rat white matter were quantitatively investigated with transmission electronic microscopy and stereological techniques. Results The white matter volume, the total length and total volume of the myelinated fibers in the white matter and the total volume of axons in the white matter of EE rats were all significantly larger than those of SE rats. The mean inner diameter and outer diameter of myelinated fibers, the total volume of myelin sheaths, the mean thickness of myelin sheaths, the mean inner perimeter and outer perimeter of myelin sheaths and the cross-sectional areas of the myelinated fibers, myelin sheaths and axons were not significantly different between EE and SE rats. Conclusion Short-term enriched environment has positive effects on the white matter and myelinated fibers in the white matter in 24-month male rats. These results suggest that short-term enriched environment might induce the remyelination of the demyelinated fibers in the white matter of aged male rats. Related Articles | Metrics

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Study on the cellular differentiation of the proliferating cells in the spinal cord of adult amyotrophic lateral sclerosis transgenic mice 2009, 40 (5):  715-719.  doi: 10.3969/j.issn.0529-1356.2009.05.005 Abstract ( )   Objective To explore the characteristic and differentiation of the proliferating cell in the spinal cord of adult amyotrophic lateral sclerosis(ALS) transgenic mice. Methods BrdU was injected at different time points during the symptomatic stage of the disease, and frozen sections were made. The cell characteristic and differentiation of the proliferating cell in the spinal cords of adult ALS transgenic mice were detected using double and triple immunofluorescence labeling technology. Results None BrdU/NeuN or BrdU/DCX double labeling cells were found in the central canal, gray matter and white matter of adult ALS transgenic mice spinal cord during the symptomatic stage of the disease. There were significantly NG2 labeling cells in the central canal, gray matter and white matter, expressing cells were detected a reduction in number during the symptomatic stage of the disease, and NG2 was expressed by most BrdU-labeled cells; Colocalization of A2B5 and BrdU was also detected in ALS mice. There were many BrdU/ GFAP double positive cells in the spinal cord of adult ALS transgenic mice, some were nestin positive, No BrdU/GFAP double labeling cells were found in wild type mice. Conclusion The neurodegenerative process stimulates a regenerative response, and there is significantly increased gliogenesis but absence of convincing neurogenesis. These data suggest that endogenous neural regeneration is insufficient for compensating the neurodegeneration. Related Articles | Metrics

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Effect of basic fibrolast growth factor on neuronal apoptosis and astroglial activation after traumatic brain injury in mice 2009, 40 (5):  720-723.  doi: 10.3969/j.issn.0529-1356.2009.05.006 Abstract ( )   Objective To study the effect of basic fibroblast growth factor (bFGF) on neuronal apoptosis and gumnosis in cerebral cortex and hippocampus tissue in mice with traumatic brain injury (TBI). Methods The mice models of TBI were established by Feeney’s method. Thirty-six mice were divided randomly into three groups including control group, saline group and bFGF group (n=12 for each group)After 3 days and 7 days，double-labeled immunoflurescence was used to detect the apoptosis of neurons. Using double-labeled immunoflurescence and Western blotting, we observed the expressions of caspase-3 in cerebral cortex and hippocampus tissue. The expression of glial fibrillary acidic protein (GFAP) in cerebral cortex was detected by Western blotting. Results The expressions of caspase-3 of cerebral cortex and hippocampus in bFGF group were lower than that in saline group (P<0.05). The expressions of GFAP of cerebral cortex in bFGF group were higher than that in saline group(EM>P/EM><0.05), there were no difference on expression of caspase-3 and GFAP between control group and saline group(EM>P/EM>>0.05). Conclusion After traumatic brain injury, bFGF could reduce the expression of caspase-3 and inhibit the neuronal apoptosis in cerebral cortex and hippocampus, increase the expressions of GFAP and promote the activation of astrcytes in cerebr Related Articles | Metrics

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Effects on the structure of cerebral temporal lobe cortex and capability of learning and memory of filial mice after administration of heroin and ephedrine 2009, 40 (5):  724-731.  doi: 10.3969/j.issn.0529-1356.2009.05.007 Abstract ( )   Objective To study effects of heroin and ephedrine on the structure of cerebral temporal lobe cortex and capability of learning and memory in filial mice. Methods One hundred and eight filial mice were given intraperitoneal injection of heroin and ephedrine by gradually increase of doses, the filial mice behavior changes were observed by bait maze. In the same time, the changes of the cell structure of cerebral temporal lobe cortex were observed by light microscope and transmission electron microscopy. Expression of Bax protein and keratinocyte growth factor(KGF) were measured by immunohistochemical method, the ChAT activity was detected by colorimetry. Results Error frequency and total test time of bait maze in the heroin group and ephedrine group were significantly higher than those in the control group(EM>P/EM><0.05 or EM>P/EM><0.01) except those at 5 days in the ephedrine group (EM>P/EM>>0.05), and the above-mentioned two indexes of the heroin group were significantly higher than those of the ephedrine group(EM>P/EM><0.05 or EM>P/EM><0.01). After administration of heroin and ephedrine at 5,10,15,20 days, the dendrites and axon of pyramidal neurons in cerebral temporal lobe cortex were atrophied and the cell bodies became smaller. Nuclear membrane lost normal round contour and the structures of organelles showed abnormal or vacuolar. The matrix around capillary dissolved or were destructed and capillary lumen became narrow. The synaptic vesicles were reduced. The number of apoptotic or necrotic cells and Bax protein and KGF-immunopositive neurons was significantly higher and ChAT activity was lower than that of the control group after administration of heroin and ephedrine(EM>P/EM><0.05 or EM>P/EM><0.01). There were differences between the heroin group and ephedrine group in the number of apoptotic or necrotic cells and expression of Bax protein and KGF(EM>P/EM><0.05 or EM>P/EM><0.01). Error frequency of bait maze, the number of apoptotic or necrotic cells and Bax proteinand KGF-immunopositive neurons were increased by the increase in doses of heroin and ephedrine. Conclusion Heroin and ephedrine had great effect on learning and memory in filial mice and this effects could be related with the damage of the histological structure of cerebral temporal lobe cortex and low activity of ChAT. Related Articles | Metrics

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Neurotoxic effects of inflammation on dopaminergic neurons in the substantia nigra in the rhesus 2009, 40 (5):  732-736.  doi: 10.3969/j.issn.0529-1356.2009.05.008 Abstract ( )   Objective To observe the neurodegenerative effects of inflammation, induced by intra-cerebral injection of lipopolysaccharide (LPS) on dopaminergic neurons in the substantia nigra of rhesus mI>onkey/I>, in order to explore the relations of mechanism of Parkinsonism with activation of microglia and the releasing of inflammatory factors in the animal brain. Methods Eight adult rhesus mI>onkey/I>s were divided into an acute experimental group and a chronic experimental group. Among them 3 mI>onkey/I>s were used for acute experiment. They were unilaterally injected with LPS and survived for 1 week to demonstrate the changes of tyrosine hydroxylase(TH) expression and inflammatory factor release by Western blotting analysis. The other 5 mI>onkey/I>s were used for chronic experiment and 4 of them were intranigrally injected with LPS and survived for 24 weeks and 48 weeks respectively after the injection. Only one mI>onkey/I> was injected with saline as a control animal. The injections were stereotaxically made 3 times(at weeks 0，8，16 respectively) in the substantia nigra on one side. The number of TH-positive neurons was detected by TH immunohistochemistry and the nigro-striatal levels of neurotransmitters were determined by high pressure liquid chromatogram (HPLC). Result In the acute experimental group of mokeys: no obvious changes of TH expressing could be seen either on the injection side and control side, but it was shown the activation of microglia and increasing expression of HLA-DR TNF-α，IL-1β and COX-2 in the substantia nigra on injected side. In the chronic experimental group of mokeys， however, the number of TH-positive neurons in the injected substantia nigra and neurotransmitters levels in the nigro-striatal area were decreased. No obvious changes could be seen in the control case of saline injected mI>onkey/I>. Conclusion LPS injection in the substantia nigra of the primate causes severe inflammation in one week, which may lead to a Related Articles | Metrics

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Action of activated microglia in hippocampal neurons of rat damage induced by hypoxia 2009, 40 (5):  737-742.  doi: 10.3969/j.issn.0529-1356.2009.05.009 Abstract ( )   Objective In order to investigate the mechanism of hypoxia damage in which the hypoxia-induced- microglia activation acts to hippocampal neurons, the indirect co-culture system was established by using the culture medium which had already trained by microglia cell N9 then used to cultivate fetal rat hippocampal neurons. Methods The co-culture system was established. The neuron growth and Caspase-3 activity of different groups were explored by TUNEL method and chemiluminescence. The expression levels of NO, OSUB>2/SUB>SUP>-/SUP> and TNF-α in different group were detected by immunofluorescence, Griess reagent method, WST-1 method and ELISA method. Results In N9 cells culture medium after 12 hours hypoxia the proliferation, vitality and apoptosis of neuron of conventional cultured and hypoxia cultured were inhibited. The stress-induced neurotoxic factors, NO, OSUB>2/SUB>SUP>-/SUP> and TNF-α in the altogether hypoxic culture system have the highest level than those in the pure culture and conventional co-culture system. Conclusion Microglia activation plays an important role in hypoxia-induced neuronal damage. The effector molecule is the neurotoxic molecule pr Related Articles | Metrics

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Polymorphism distribution of 15 STR in Lasha and Naqu tibetan population 2009, 40 (5):  743-748.  doi: 10.3969/j.issn.0529-1356.2009.05.010 Abstract ( )   Objective To investigate the genotype and allele frequency distribution of 15 STR loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818 and FGA)in 408 Lasha and Naqu Tibetans. Methods Extracting genome DNA, then multiplex PCR amplification using five fluorochromes (6FAM, VIC, NED, PET, LIZ) were used to acquire the genotype frequency and allele frequency ,and all loci were tested for HardyWeinberg equilibrium, at the same time to compare the allele frequency between Lasha and Naqu Tibetans. Results In Lhasa Tibetan population, 11-47 genotypes and 5-12 alleles were detected in 15 STRs. In Naqu tibetan population, 12-58 genotypes and 6-14 alleles. Allele frequency had little difference between Lasha and Naqu Tibetan population. The gene frequency of these 15 STR loci were in Hardy-Weinberg equilibrium.Conclusion These 15 STR loci have highly polymorphism, and the 15 STR loci could be used as the geneitc markers of Tibetan populations in anthropological studies, linkage analysis of genetic diseases, individual identification and paternity testing in forensic medicine. Related Articles | Metrics

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Recombinant BJ46a protein exerts inhibition effect on invasion and metatasis of B16 cells EM>in vitro/EM> 2009, 40 (5):  749-754.  doi: 10.3969/j.issn.0529-1356.2009.05.011 Abstract ( )   P>Objective To investigate the effects of BJ46a on invasion and metastasis of melanoma cells. Methods A snake venom metalloproteinase inhibitor BJ46a was produced in baculovirus expression system. Then, recombinant fusion protein was purified by ProBond at the point of maximal expression, SDS-PAGE and Western blotting were used to detect the protein expression. The inhibition activity of matrix metalloproteinasel(MMPs) was detected by purified BJ46a protein. The abilities of tumor cell proliferation and adhension were determined by Alamar blue assay(EM>n/EM>=3).The ability of tumor cell invasion was identified by transwell chamber EM>in vitro/EM>(EM>n/EM>=3). Results The activity of MMPs could be reduced by purified BJ46a protein. The ability of B16 cells treated with recombinant BJ46a to invade the reconstituted basement membrane decreased significantly(EM>P/EM><0.01)，inhibition ratio was 84.8%, but the proliferation and adhension abilities had no change.Conclusion BJ46a might has a strong inhibition effect on the invasion and metatasis potential of B16 cells EM>in vitro/EM>./P> Related Articles | Metrics

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Regulation of progesterone on interleukins and TNF-α expression ofmurine CD11c positive splenic dendritic cells 2009, 40 (5):  755-759.  doi: 10.3969/j.issn.0529-1356.2009.05.012 Abstract ( )   Objective To elucidate the effect of progesterone (Prog) on the cytokine expressions of murine CD11c positive splenic dendritic cells (SDCs). Methods SDCs were purified by immune beads and treated with various concentrations of Prog for 24hours, then cytokine expressions of SDCs in the mRNA level were analyzed through reverse transcription-polymerase chain reaction (RT-PCR). Results Prog increased the transcript of IL-6 in middle dose group but decreased it in high dose, and the level of IL-10 was increased in low and middle dose groups while decreased in high dose group. Furthermore, the transcripts of IL-12 and TNF-α were increased and decreased respectively in all Prog treated groups.Conclusion Prog might regulate organ immune response through influencing cytokine expressions of SDCs. Low- and middle-dose Prog switch organ immune response to Th2-type, while high-dose Prog develops a Th1-type immune response. Related Articles | Metrics

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Expression of stanniocalcin-1 mRNA in rat hepatocarcinogenesis induced by diethylnitrosamine 2009, 40 (5):  760-765.  doi: 10.3969/j.issn.0529-1356.2009.05.013 Abstract ( )   P>Objective In the present study, levels of STC1 mRNA and STC1 were detected at various stages in rat hepatocarcinogenesis induced by diethylnitrosamine (DENA). Methods Eighty SpragueDawley rats were divided into experimental group and the control group. In the experimental rats, the hepatocarcinogenesis was induced with diethylnitrosamine (DENA) through intragastric gavage at a dose of 70 mg/kg body weight, and the control rats were done with distilled water at the same dosage. The cellular changes in liver were determined with histopathological techniques. Both of STC1 mRNA and STC1 level in liver were detected with RT-PCR and Western blotting technique, respectively. Results The results showed that both of STC1 mRNA and STC1 were only detected in liver of rats exposured to DENA. Conclusion The data suggested that there was correlation between the expression of STC1 gene and hepatocarcinogenesis, and implied STC1 as potential novel prognostic marker for detecting human liver cancer. Mammalian stanniocalcin1 (STC1) is a glycoprotein hormone expressed in abroad spectrum of tissues such as kidney, prostate, ovary, etc. Its expression is found to be regulated in numerous developmental, physiological and pathological processes, including pregnancy, lactation, cancer, apoptosis, organogenesis, etc./P> Related Articles | Metrics

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P>Effect of estrogen replacement threapy on Cx43 expression and metal elements in ovariectomy and chronic aluminum toxication rat heart /P> 2009, 40 (5):  766-769.  doi: 10.3969/j.issn.0529-1356.2009.05.014 Abstract ( )   Objective To reveal the influence of estrogen replacement threapy on the gap junction connexin 43(Cx43)expression and content of metal elements in ovariectomy and chronic aluminum toxication rat heart. Methods Forty 6-month-old female SD rats were divided randomly into 4 groups: group Sham, group OVX, group OVX+Al and group OVX+Al+ESUB>2/SUB>. After 3 months, The content of metalelements in heart was detected by ICP-AES. The Cx43 expressions were observed and determinated by immunohistochemistry and digital image analysis. Results 1.Cx43 positive granules distributed in cell side surface and cytoplasm as well as intercalated disk in atrial myocytes. They typically confined to the site of intercalated disk in ventricular myocytes in group Sham. In Group OVX, the Cx43 of atrial myocytes were redistributed from cell side surface to the intercalated disk. They were redistributed from the intercalated disk to cell side surface and in cytoplasm in ventricular myocytes. And some Cx43 granules distruibuted irregularly. The Cx43 redistribution of group OVX+Al was more irregular than that of group OVX. In group OVX+Al+E2, the Cx43 distribution of atrial myocytes returned to normal. 2. The amount of Cx43 in all groups was not changed. 3. In sham group，the content of mental elements in heart showed as follows: Mg＞Ca＞Fe＞Zn＞Al＞Si＞Cu＞Mn＞Se and Cd; Compared between group OVX and group sham, the content of Zn decreased (EM>P/EM><0.001); Compared between group OVX+A1+ESUB>2/SUB> and group sham, the content of Al, Cd, Si and Se increased (P<0.05，EM>P/EM><0.01，EM>P/EM><0.01，EM>P/EM><0.001); Compared between group OVX+Al+ESUB>2/SUB> and group OVX+Al, the content of Cd, Mn and Se inc Related Articles | Metrics

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Pentoxifylline have anti-inflammatory effect on nonalcoholic steatohepatitis in rats 2009, 40 (5):  770-775.  doi: 10.3969/j.issn.0529-1356.2009.05.015 Abstract ( )   Objective To study the effects of pentoxifylline (PTX) on antiinflammatory factor of nonalcoholic steatohepatitis (NASH) in rats. Methods Rats fed a standard diet for one week were randomly divided into four groups, control group(EM>n/EM>=10),12 weeks model group(n=10), 16 weeks model group(EM>n/EM>=10) and the PTX treatment group(EM>n/EM>=10). Two model groups and the PTX treatment group were then fed a high-fat diet. Meanwhile the rats fed a standard diet served as control group. After 12 weeks, the 12 weeks model group were sacrificed and the treatment group were given PTX ［16mg/(kgI>&#/I>8226;d)］ by intragastric administration. Then all the rats of other three groups were sacrificed at the end of the 16th week. Blood samples were collected aseptically from the abdominalis aorta and serum was separated with regular methods. Serum level of tumour necrosis factor α(TNF-α)was determined by ELISA. Serum levels of IL-1β and IL-6 were tested by radio immunoassay(RI). The immunohistochemical staining was perfored on the liver tissue slice, the expressions of IL-1β, IL-6 were evaluated. TNF-α mRNA was detected by RT-PCR. NF-κB binding activity was detected by electrophoretic mobility shift assay(EMSA). Results Serum level of TNF-α, IL-1βand IL-6 of model groups increased significantly than that of the control group. No statistical difference was found among the groups. The expressions of IL-1β，IL-6 were increased significantly in the model groups and the PTX treatment group (vs. the control group, P<0.05), while the expression of IL-6 was decreased in the PTX treatment group (vs. the model groups); The level of TNF-α mRNA was increased significantly in the 16 weeks model groups (vs. the co Related Articles | Metrics

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Study on the cell morphology of glucocorticoid-induced murine thymocyte death 2009, 40 (5):  776-779.  doi: 10.3969/j.issn.0529-1356.2009.05.016 Abstract ( )   Objective To study cell morphological change of glucocorticoid(GC)induced murine thymocyte death. Method After intraperitoneal injection of dexamethasone into 4-6-week-old female BALB/C mice, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end-labeling (TUNEL) method was used to detect the DNA fragmentation or double strand breaks. To examine acid phosphatase (ACP), we applied double staining of the section with the TUNEL method and ACP. Moreover, we used transmission electron microscopy to observe ultrastructure of apoptosis cells and macrophages as well as the activity of macrophages after apoptosis. Results We demonstrated under light microscopy that, the TUNEL positive thymocytes were scattered throughout the cortex of GC-treated thymus, and the apoptosis index was 0.460±0.012;In the control group, the TUNEL positive ones were few and the apoptosis index was 0.020±0.001. There was significant difference between the two groups(EM>P/EM><0.05). Double staining revealed that all the TUNEL positive cells were phagocytosed by ACP positive macrophages. It was also observed by transmission electron microscopy, and these small unphagocytosed thymocytes were apparently dead cells, as based on the extent of chromatin condensation was enormous. Conclusion Typical apoptosis, which is characterized by DNA fragmentation, was not the dominant type of GC-induced murine thymocyte death. These dead thymocytes were phagocytosed by ACP positive cells.The most important effect of GC for thymocyte was that it can induce chromatin pycnosis without DNA fra Related Articles | Metrics

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Expression of c-Met in muscle satellite cells of muscle injury 2009, 40 (5):  780-784.  doi: 10.3969/j.issn.0529-1356.2009.05.017 Abstract ( )   Objective To observe the expression of c-Met in muscle satellite cellsEM>in situ/EM> of normal and cardiotoxinintoxicated muscle tissues. Methods Twelve CSUB>57/SUB> mice were divided into six groups, and raised in IVC laboratory. In the 1st to 6th group, the mice were injectied with cardiotoxin (5μg per mouse) in left quadriceps femoris, while their right quadriceps femoris were left for control without any injection.After the mice were sacrificed at different limited timepoint. (one day, four days, one week, two weeks, four weeks, six weeks), the quadriceps femoris were isolated and fixed in 4 percent formaldehyde for 24 hours and then embeded in paraffin for histological, immunohistochemistical and reverse transcriptionpolymerase chain reaction (RT-PCR) analysis to detect the expression of c-Met in muscle tissues EM>in situ/EM>. Results The results of immunohistochemistry and RT-PCR showed that there were few expressions of c-Met in muscle satellite cells of normal muscle tissues. Compared with normal tissues, obvious expression was found in 24 hourgroup as well as the sixth weekgroup. The expression of c-Met peek Related Articles | Metrics

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Localization and possible roles of the Ran protein in rat skin fibroblasts 2009, 40 (5):  785-788.  doi: 10.3969/j.issn.0529-1356.2009.05.018 Abstract ( )   Objective To study the localization of the Rasrelated nuclear protein Ran in rat somatic cells, and to investigate its interaction with the protein associated to the cell cycle. To reveal the molecular mechanism of the cell cycle control and mitotic division. Methods Fibroblasts derived from rat skin were successfully cultured with a normal tissue culture method，and investigated the localitation and possible roles of Ran during the cell mitotic division by using laser scanning confocal microscopy, repeated three times at least. Results The results showed that Ran was localized mainly in the nucleus, except for the nucleolus, in the intermediate stage rat skin fibroblasts cell. Its density was very low in the cytoplasm. After the nuclear membrane broke down in mitochysis, Ran distributed throughout the cell, but concentrated to the mitotic spindle, the concentrated area was uniformity with the spindle; in anaphase and telephase, the concentrated area changed with the spindle, localized between the separated chromosome, but lower in other sites. After mitosis, Ran migrated into the forming nucleus. There was no special distribution for Ran under the membrane of the cell fission. Conclusion Ran has a cell cycle-dependent localization and this is consistent with its regulatory roles in nuclear import and export in rat intermediate cells, the spindle assembly correctly, the correct chromosome segregation, and the cell cycle regulation. Related Articles | Metrics

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Inducing mouse mesenchymal stem cells to differentiate into insulin_producing cells by rat pancreatic extract EM>in vitro/EM> 2009, 40 (5):  789-793.  doi: 10.3969/j.issn.0529-1356.2009.05.019 Abstract ( )   Objective To research the method of mice bone marrow mesenchymal stem cells (BMMSCs) trans-differenting into insulin producing cells (IPCs) by rat pancreatic extract (RPE). Methods Mesenchymal stem cells were isolated from Kunming mouse, and RPE was from SD rats’ pancreases. Passaged BMMSCs were induced to be insulin-producing cells (IPCs) with RPE. First, devided mice’s BMMSCs to 6 groups,3 culture flasks per group at random, and added RPE 10mg/L, 20mg/L, 30mg/L, 40mg/L, 50mg/L, 100mg/L to each group. After 1 week, decided which concentration of RPE was better. And induce mouse BMMSCs to IPCs by it (50mg/L). The expression of insulin in IPCs was detected by immunoenzyme cytochemistry and immunofluorescence cytochemistry, the trans-differentiation purity of IPCs was detected by dithizone (DTZ)stain，C-peptide secreting was detected by radioimmunoassay(RIA). Results Mesenchymal stem cells of mice could be induced into IPCs by 50mg/L PRE. Typical islet-like clustered cells were observed after 1weeks, and could be stained scarlet by DTZ.The insulin expression of BMMSCs was positive by immunoenzyme cytochemistry and immunofluorescence cytochemistry. RIA showed the expression of C-peptide in the IPCs could be detected. Conc Related Articles | Metrics

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Effect of osteopontin on mouse blastocyst adhesion and outgrowth and its mechanism 2009, 40 (5):  794-797.  doi: 10.3969/j.issn.0529-1356.2009.05.020 Abstract ( )   Objective To investigate the effect of osteopontin on mouse blastocyst attachment and outgrowth EM>in vitro/EM> and its mechanism. Methods Mouse blastocysts were cultured in microdrops,and the culture plates were precoated with fibronectin. Blastocysts were cultured EM>in vitro/EM> in Quinn’s medium containing various concentrations of recombinant mouse osteopontin, osteopontin antibody and RGD peptide.The percent of hatched blastocysts,blastocysts with adhesion and outgrowth were calculated at 24 hours,48 hours and 72 hours after culture. In another experiment,blastocysts were cultured in 24-well culture plate, the concentrations of matrix metalloproteinases (MMP)-2, -9 of culture medium were detected by ELISA. Results There were no significant difference in percent of hatching and attachment between control and OPN treated groups, however, the rates of outgrowth in OPN treated groups at concentrations of either 1.0mg/L or 10.0mg/L were higher than those in control group after 72 hours culturing(EM>P/EM><0.05 and EM>P/EM><0.01,respectively),and the blastocysts started to outgrowth in advance after 48 hours culturing, whereas mouse osteopontin antibody and RGD reduced the incidence of blastocyst hatching, attachment and outgrowth significantly in a dose-dependent manner. Osteopontin promoted the blastocysts secreting MMP-2, -9 in a dose-dependent and time-dependent manner.Conclusion Osteopontin is involved in regulating embryo early implantation process EM>in vitro/EM> by promoting blastocyst outgrowth and secreting MMP-2, -9. Related Articles | Metrics

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Research on histological structure of autofluorescence of EM>Dugesia japonica/EM> 2009, 40 (5):  798-802.  doi: 10.3969/j.issn.0529-1356.2009.05.021 Abstract ( )   Objective Hermaphroditic planarians possess a very important position in the systematic evolutionary history of animal, as they are capacity of complete regeneration. Hence, the research on histological structure of autofluorescence has been carried out to provide a crucial insight into the developmental and regenerative biology. Methods Part of histological structure of planarian (EM>Dugesia japonica/EM>) was revealed with HE method, Masson method and Van Gieson method. Their autofluorescence was observed with ultraviolet. There were six planarians in each stained group and the autofluorescence group. Results Epidermis, outer epidermis of pharynx, protonephridium, intestine, the photoreceptor cells and longitudinal nerve cords, all radiated blue autofluorescence. The epithelial dissociation side of copulatory bursa radiated yellow autofluorescence, its middle part radiated blue autofluorescence, its fundus side radiated weakly blue autofluorescence. Testis could hardly give off autofluorescence. Pigment cells of eyepot could not give off autofluorescence. Conclusion The research on configuration and autofluorescence of planarian eye may offer help for the study of origin and evolutionary law on eye of invertebrate. Related Articles | Metrics

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Effect of neonate rats’ cardiomyocytes transplantation on myocardial capillary density of rabbits with acute myocardial infarction 2009, 40 (5):  803-806.  doi: 10.3969/j.issn.0529-1356.2009.05.022 Abstract ( )   Objective To explore the influence of neonate rats’ cardiomyocytes transplantation on myocardial capillary density and microvessel regeneration of rabbits with acute myocardial infarction(AMI). Methods Sixteen New Zealand white rabbits and 40 neonate rats were used in the study. After the model of AMI in rabbits was established, cultured neonate rats’ cardiomyocytes were injected into the area of AMI. Specimens were collected 1, 2, 3 and 4 weeks after transplantation, and they were stained by alkaline phosphatase(AlP) histochemical method and immunofluorescence method, then observed by light microscopy. The capillary density and regeneration around the marginal zone of AMI were assayed through LEICA Qwin V3 image analysis system. Results After neonate rats’ cardiomyocytes transplantation for 1, 2, 3 and 4 weeks, the capillary density and the number of regenerate capillary around the area of rabbit’s AMI were increased significantly than those of the control group. And through quantitative analysis, we found that myocardial capillary density tend to be increased first and then decreased.Conclusion Allogenic cardiomyocytes transplantation could promote neovascularization of acute infarcted myocardium of rabbits, and then improve the Related Articles | Metrics

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Effect of 17β-estradiol on myocardial ischemia-reperfusion injury of rats 2009, 40 (5):  807-810.  doi: 10.3969/j.issn.0529-1356.2009.05.023 Abstract ( )   Objective To explore effects of 17β-estradiol preconditioning on myocardial ischemia-reperfusion(I/R) injury of rats and its mechanism. Methods Totally 30 female SD rats were ovariectomized and divided randomly into C group (sham), I/R group and I/R+ESUB>2 /SUB>group (17β-estradiol pretreatment). After 2 weeks of pretreatment with 17β-estradiol, all rats were subjected to the left anterior descending coronary occlusion for 30 minutes and reperfusion for 2 hours. The sizes of myocardial infarction area were measured by triphenyltetrazolium chloride (TTC) staining; the apoptotic rate was assayed by flow cytometer, the protein expression of EM>c-fos/EM> proto-oncogene in cardiomyocytes was detected by semiquantitative immunohistochemical technique. Results 17β-estradiol significantly attenuated the infarct size of myocardium injured by ischemia-reperfusion, decreased the apoptotic rate in cardiomyocytes, also inhibited the expression of c-Fos protein in cardiomyocytes.Conclusion These results suggest that estrogen protects the heart against ischemia-reperfusion injury of rat，its cardioprotective mechanism Related Articles | Metrics

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Effect of angiotensin Ⅱ on cell proliferation and apoptosis in developing renal corpuscles of mouse 2009, 40 (5):  811-815.  doi: 10.3969/j.issn.0529-1356.2009.05.024 Abstract ( )   Objective To observe effects of angiotensin Ⅱ(AngⅡ) on proliferation and apoptosis of developing mouse renal corpuscle, derived from the 12-day embryo EM>in vitro/EM>. Methods Totally 40 pregant female mice(embryonic 12 days)were randomly divided into six groups: control group and groups which were induced by different concentrations of Ang Ⅱ(10SUP>-9/SUP>-10SUP>-5/SUP>mol/L). The kidney tissues were cultured for 2，4，6 days. Immunohistochemistry(PCNA and TUNEL),double immunofluorescence and laser scanning confocal micropcopy were used to observe the expressions of PCNA and TUNEL positive cells in mouse renal corpuscles at different developmental stages. Results AngⅡ was able to induce the renal corpuscular proliferation and apoptosis to a certain degree,and the effects were in accordance with AngⅡ concentration and inducing time. The shorter AngⅡ-induced time or the lower concentration of AngⅡ was the more effective on the proliferation, on the contrary, the longer AngⅡ-induced time or the higher concentration of AngⅡ, the more significant effect on the apoptos Related Articles | Metrics

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Temporal-spatial expression of SHH and β-catenin during hair follicle morphogenesis in different hoof periphery site of EM>Bos grunniens/EM> embryos 2009, 40 (5):  816-820.  doi: 10.3969/j.issn.0529-1356.2009.05.025 Abstract ( )   Objective To investigate hair follicle morphogenesis of different hoof periphery site, SHH and β-catenin temprocal-spatial expression during hair follicle morphogenesis. Methods Totally 41 embryonic 45-248 days Yak embryos forepaw were used as material to investigate hair follicle morphogenesis of different hoof periphery site by HE staining and immunohistochemistry of tissue section for detecting temprocal-spatial expression and epithelial-mesenchymal reciprocal interaction during morphogenesis. SHH expression during hair follicle morphogenesis showed stronger effect on the hair follicle epidermis than the hair follicle mesenchyme. Results The hair follicle morphogenesis of yak proximal hoof end and distal hoof end initiate from embryonic 45 days and initiate from embryonic 55 days respectively, mature hair follicle in embryonic 120 days. From the proximal hoof end to distal hoof end, the time,the phase,the size and the growing depth of the hair follicle morphogenesis in turn showed a occure degressive gradient. β-catenin expression during hair follicle morphogenesis showed a stronger effect on the hair follicle epidermis than the hair follicle mesenchyme.Conclusion We speculate that SHH and β-catenin expression and detail function difference play important roles during hair follicle morphogenesis in different hoof periphery site of yak embryos. The epidermis is dominated during the hair follicle mor Related Articles | Metrics

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Histological characterization of pulmonary alveoli of one-day old yak 2009, 40 (5):  821-825.  doi: 10.3969/j.issn.0529-1356.2009.05.026 Abstract ( )   Objective To find out the characteristics of pulmonary alveoli histological structure and the adaptive mechanisms of 1-day old yak and its relations to the plateau hypoxia environment. Methods To analyze pulmonary alveoli of five 1-day old yaks and cattle respectively by HE staining, transmission electron microscopy(TEM) and morphometry. Results Pulmonary alveoli of 1-day old yak dilated sufficiently more than that of the cattle. There were massive sac-like structures which resemble alveolar sac in the lung of 1-day old yak. There was large bud-like septum in each saclike structure. The saclike structure was rare in the 1-day old cattle’s lung. Type Ⅱ alveolar cells were abundant in the 1-day old yak alveolar wall, and lamellar bodies were obvious in the yak alveolar cavity. Mean alveolar number(MAN) of 1-day old yak is less than that of the 1-day old cattle’s(EM>P/EM><0.05);but mean single alveolar area (MSAA), alveolar area in the unit area (SA) and mean thickness of alveolar septum(MSAT)in the yak alveoli were higher than cattle’s corresponding values(EM>P/EM><0.05). Arithmetic mean thickness of 1-day old yak blood-air barrier was thicker significantly than that of the 1-day old cattle’s(EM>P/EM><0.05)，but harmonic mean thickness of blood-air barrier was not significant (EM>P/EM>>0.05). Conclusion The development and growth of alveoli of 1-day old yak is more mature than that of the cattle’s. Yak alveoli increase rapidly by way of bud growth. Comparatively mature development and growth of lung，rapid increase in Related Articles | Metrics

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Expressions of 14-3-3 ζ and β-catenin proteins in human TSUB>1/SUB> non-small cell lung cancer and the significance 2009, 40 (5):  826-829.  doi: 10.3969/j.issn.0529-1356.2009.05.027 Abstract ( )   Objective To observe the expressions of 14-3-3 ζ and β-catenin in human TSUB>1/SUB> nonsmall cell lung cancer(NSCLC), and to investigate the relationship between the expressions of 14-3-3 ζ and β-catenin and progression and invasion in NSCLC. Methods The expressions of 14-3-3 ζ and β-catenin were detected by immunohistochemistry, double fluorescent, laser scanning confocal microscopy and Western blotting in NSCLC and normal lung. Results Upregulation of 14-3-3 ζ was observed significantly in stage TSUB>1/SUB> NSCLC in contrast to that in normal lung. The increased 14-3-3 ζ protein level was correlated with differentiation degree and lymph node metastasis, but it was no correlation with histological type. Decreased expression of β-catenin was observed in NSCLC in contrast to that in normal lung. The expression of β-catenin was positively related to differentiation degree and was negatively related to the lymph node metastasis, and there was no relationship between β-catenin expression and histological type. Conclusion 14-3-3 ζ a Related Articles | Metrics

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Expression of forkhead/ winged helix transcription factor in the liver of BXSB mice 2009, 40 (5):  830-833.  doi: 10.3969/j.issn.0529-1356.2009.05.028 Abstract ( )   Objective To present gene and protein expressions of forkhead/winged helix transcription factor (Foxp3) in the liver of BXSB mice and to explore potential roles of regulatory T cell(Treg) in pathogenesis of hepatic lesions in BXSB mice. Methods We examined the liver expression of Foxp3 in 8-and 16-week BXSB male mice using immunohistochemical staining and real-time PCR, and compared with 8-week normal C57BL/6 male mice as a control. Results In immunohistochemistry，Foxp3 staining in normal control mice was pretty strong as dark brown and distributed in hepatic sinusoid and perisinusoidal space, but in 8-and 16-week BXSB mice the staining was weaker as light brown than normal mice and distributed near limiting plate mainly. The mRNA of Foxp3 was significantly down-regulated in 8-and 16-week BXSB mice liver compared to normal controls(0.30±0.04，0.18±0.03 vs 1.08±0.08,EM>P/EM><0.01), and the Foxp3 expression of 16-week BXSB mice was furthermore downregulated than that of 8-week BXSB mice(EM>P/EM><0.01);Conclusions Distribution and down-regulation of Foxp3 in Treg may play a role in t Related Articles | Metrics

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Comparative study between the sectional anatomy and MRI of the interosseous talocalcaneal ligament and its clinic significance 2009, 40 (5):  834-836.  doi: 10.3969/j.issn.0529-1356.2009.05.029 Abstract ( )   Objective To provide sectional anatomical basis for clinical magnetic resonance imaging (MRI) diagnosis of the interosseous talocalcaneal ligament. Methods Respectively,twelve cadaveric right feet taped in 20°plantarflexion position were frozen and cut into 3-mm-thick slices in coronal(EM>n/EM>=4),sagittal(EM>n/EM>=4) and transverse(EM>n/EM>=4) planes.Then we compared the slices with the MR images. Results In sagittal section through lateral border of collum tali, interosseous talocalcaneal ligament is within (19.2±2.8)mm. In coronal section through medial part of corpus tali, interosseous talocalcaneal ligament is within (17.9±5.2)mm.In the sagittal section through medial part of corpus tail, interosseous talocalcaneal ligament can be displayed most distinctly and it is 17.2mm long,11.9mm wide in sagittal diameter. In coronal section through medial part of corpus tali, interosseous talocalcaneal ligament can be displayed most distinctly and it is 13.1mm long,14.1mm wide in transverse diameter. In transverse section,the interosseous talocalcaneal ligament can not be displayed clearly.Conclusion MRI provides excellent delineation of the interosseous talocalcaneal ligament and evidence for the diagnosis of the interosseous talocalcaneal ligame Related Articles | Metrics

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Expressions of nNOS and VEGF in the early developing small intestinal villi of human fetus and its significance 2009, 40 (5):  837-839.  doi: 10.3969/j.issn.0529-1356.2009.05.030 Abstract ( )   Objective To explore the rule of the expressions of neuronal nitric oxide synthase(nNOS) and vascular endothelial growth factor(VEGF) in the early developing human fetus. Methods At the second to fourth month of gestation, the expressions of nNOS and VEGF were observed in 14 cases of human fetal small intestine villi by using the SABC and PV of the immunohistochemical methods. Results In the second month of gestation, the expression of nNOS was negative and VEGF was positive at the epithelial cell layer in the small intestinal villi of human fetus. At the third to fourth month of gestation, the expressions of nNOS and VEGF proteins were positive in some cells of the small intestinal villi of human fetus. Conclusion nNOS and VEGF proteins are closely related to the growth and development of the cells and tissues in the early developing small intestinal villi of human fetus. Related Articles | Metrics

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Comparison between two methods for displaying microvessels in rat different tissues 2009, 40 (5):  840-843.  doi: 10.3969/j.issn.0529-1356.2009.05.031 Abstract ( )   bjective To investigate the differences between CD34 immunohistochemistry and NBT/BCIP enzymic histochemistry in displaying microvessels of various tissues. Methods The microvessels in rat small intestinal, kidney, spleen, liver, heart, lung and skin were stained by CD34 immunohistochemistry and NBT/BCIP enzymic histochemistry, and the two methods were compared by morphological analysis and microscopic count. Results We found that the microvessels in all selected tissues above mentioned were stained by CD34 immunohistochemistry, and that the microvessels in small intestinal, kidney, liver and heart tissues, but not in spleen, lung and skin tissues, were stained by NBT/BCIP enzymic histochemistry. And in the same sections of one tissue, the microvascular density by NBT/BCIP enzymic histochemistry was higher than that by CD34 immunohistochemistry. Conclusion CD34 immunohistochemistry is generally suitable to display the microvesssels in various organs, while NBT/BCIP enzymic histochemistry shows some organ microvessels at higher density. Related Articles | Metrics

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Morphological features and functional implications of autophagy 2009, 40 (5):  844-849.  doi: 10.3969/j.issn.0529-1356.2009.05.032 Abstract ( )   Objective Autophagy is a process by which cells phogocytize intracellular cytoplasm or organelles when cells are stimulated and cargoes are eventually degraded in lysosomes. According to the ways in which cargoes are transported into lysosomes, types of autophagy are divided into macroautophagy, microautophagy and chaperone-mediated autophagy. In macroautophagy, the autophagosome precursor wraps cytoplasm or organelles to form an autophagosome. Then the autophagosome fuses with the lysosome forming the autophagolysosome. The cargoes are degraded in lysosome. In microautophagy, the lysosome engulfs cytoplasm, organelles or nucleus directly through invagination of the lysosomal membrane and formation of the autophagosome. Finally, the contents of the autophagosome are digested by lysosomal enzymes. In chaperonemediated autophagy, cytosolic proteins are translocated into the lysosome by lysosomal membrane receptor. Autophgy is evolutionarily conserved from yeasts to mammalian cells and plays important roles in tolerating starvation and ischemia, cleaning the senescent organelles, eliminating bacteria and foreign matters, maintaining cellular activities and extending longevity. Autophagic activities are regulated by autophagic genes. Deletion or dysfunction of autophagic genes may result in some diseases. Elucidating the processes of autophagy, autophagic structures and their functions is helpful for exploring mechanisms of roles of autophagy in physiological and pathological processes. This review focuses on morphological characteristics of the autophagic structures and functional implications of these structures. Related Articles | Metrics