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2010, Volume 41 Issue 6
06 December 2010 Previous Issue    Next Issue
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Comparison of differential proteomics between neural stem cells and motor neurons in embryonic spinal cord in rats 2010, 41 (6):  779-784.  doi: 10.3969/j.issn.0529-1356.2010.06.001 Abstract ( )   Objective To analysis and compare the proteomic differences between neural stem cells and motor neurons in embryonic spinal cord in rat and discover the key different proteins. Methods Separating the protein of cells by the 2-D fluorescence difference gel electrophoresis, and to analyse the differences of protein expression with DeCyder software, and to identify with high performance liquid chromatography-electrospray tandem mass spectrometry. Results About 1 300 protein spots from the cells were gained after gel analysis. Eighty-seven protein spots were selected for mass spectrometry analysis. Fourty-four differently expressed proteins (24 in neural stem cells and 20 in motor neurons) were identified by mass spectrometry analysis. Conclusion Differently expressed proteins between neural stem cells and motor neurons were identified and it is helpful to find the new targets in neural stem cells differentiation into motor neurons. Related Articles | Metrics

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Comparison of the differentiation capability between human parthenogenetic embryonic stem cells and normal embryonic stem cells 2010, 41 (6):  785-789.  doi: 10.3969/j.issn.0529-1356.2010.06.002 Abstract ( )   Objective The differentiation capability of human parthenogenetic embryonic stem cells (pESCs) and normal embryonic stem cells (nESCs) was compared to estimate whether the pluripotent potential of pESCs was equal to that of nESCs. Methods pESCs and nESCs were injected into the rear leg of mice with severe combined immunodeficiency disease (SCID) to form teratoma, which was then processed for histological analysis by HE staining; pESCs and nESCs were detached to grow as aggregates in suspension to form embryoid body (EB), which were collected for the analysis of the key genes expression related with the development of main organs from all three germ layers and trophoblast using RT-PCR. Further, pESCs and nESCs were induced to differentiate into trophoblasts and detected the human chorionic gonadotrophin-β(hCG-β) content in the culture medium by ELISA as well as the percentage of hCG-β positive cells by flow cytometry. Results pESCs and nESCs can differentiate into the cell types from three germ layers both EM>in vivo/EM> andEM>in vitro/EM> differentiation assay. pESCs and nESCs were able to form teratomas with a complex pattern of differentiation to three germ layers, such as neural epithelia, cartilage and glandular epithelium. The key genes related with the development of main organs from all three germ layers and tropoblast were expressed in day 5 and day 21 EB from pESCs and nESCs. pESCs can be induced to differentiate into trophoblasts and expressed hCG-β, but the content of hCG-β and the percentage of hCG-β positive cells were lower in pESCs than that in nESCs. Conclusion pESCs can differentiate into three germ layers and tropoblas Related Articles | Metrics

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Temporal expression of GABASUB>A/SUB> receptor subunits α2 and α3 in the contralateral cortex of rats after focal brain ischemia 2010, 41 (6):  790-795.  doi: 10.3969/j.issn.0529-1356.2010.06.003 Abstract ( )   Objective To study the expressions of GABASUB>A/SUB> receptor subunits α2 and α3 after middle cerebral artery occlusion/reperfusion in rats. Methods The animals in the experimental groups were subjected to transient middle cerebral artery occlusion(MCAO) using the intraluminal thread model. The rats were randomly distributed into MCAO group （EM>n/EM>=18）and sham group（EM>n/EM>=18）. Rats were sacrificed at the 7th day, 30th day and 6th month after operation. Immunohistochemical staining was used to detect the expressions of GABASUB>A /SUB>receptor subunits α2 and α3 between MCAO and sham group. Data were analyzed by SPSS software. Results In the contralateral cortex, there were no significant difference of subunits α2 and α3 expression between MCAO group and sham group in 7 days after ischemia. In 30 days and 6 months, the expression of subunit α3 in Hl,Fr,Par1 and Par2 brain regions was markedly higher than that of sham group in contralateral cortex (P <0.05). In 30 days, the expression of subunit α2 in MCAO group was higher than that of sham group in Par1 and Par2 brain regions (SUP>P/SUP> <0.05); however, in 6 months the expression of subunit α2 was lower in Par2 and higher in Par1 compared with sham group (EM>P/EM> <0.05). Conclusion The expressions of subunits α2 and α3 have a generalized and lon Related Articles | Metrics

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Prenatal alcohol exposure induces the autophagy in cerebellar Purkinje cells of mice 2010, 41 (6):  796-804.  doi: 10.3969/j.issn.0529-1356.2010.06.004 Abstract ( )   Objective To investigate the effect of ethanol exposure on the autophagy in cerebellar Purkinje cells. Methods With intubating pregnant mice ethanol daily from E5 until the pup’s birth, the animal models of prenatal alcohol exposure (PAE) were made. Totally 180 pups included control treatment and ethyl alcohol(EtOH) treatment were used in the study. On the other hand, the autophagy in cerebellar Purkinje cells in P7, P14 and P30 was investigated with transmission electron microscopy, immunofluorescent labeling and Western blotting technique. Results With electron microscopy, a numerous lysosome and autophagosomes were found in the Purkinje cells of alcohol exposure mice, with organelle injuries as well. Under light microscope, Purkinje cells changed lots after ethanol exposure, such as irregular arrangement, reduced dendritic branches and dentritic spines. The positive autophagy cells (PAC) and positive autophagy index (PAI) in ethanol treatment groups were obviously higher than that in control at either P7 or P14 (EM>P/EM><0.001) with dose dependency (EM>P/EM><0.001). In the meantime, PAC and PAI increased gradually with age increasing EM>(F/EM>=58.29,EM> P/EM><0.001). The coexpression of Beclin-1 and MAPLC3 in the same Purkinje cell suggested the Beclin-1’s regulation to the autophagy. Western blotting analysis supported the above results from immunocytochemistrys. Conclusion Autophagy is involved in the normal development and alcohol’s toxicological effects of cerebellar Purkinje cells. The prenatal ethanol exposure can induce the autophagy of Purkinje c Related Articles | Metrics

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Effect of triptolide on expression of C1q in the hippocampus of Alzheimer’s disease rats 2010, 41 (6):  805-808.  doi: 10.3969/j.issn.0529-1356.2010.06.005 Abstract ( )   Objective To investigate the effect of triptolide(TP) on expression of C1q in the hippocampus of model rats with Alzheimer’s disease(AD). Methods Thirty male SD rats were randomly divided into control group, AD model group and TPtreated group.Each group had 10 rats. The AD model group was established with bilateral microinjection of aggregated beta-amyloid protein(Aβ)SUB>1-40/SUB> into hippocampus in rats and Morris water maze test was used to judge that the model was successful or not. The control group was established with bilateral microinjection of normal saline into hippocampus in rats.The TPreated group rats were administered TP ［0.4mg/(kg.d)］ intraperitoneally after the AD model was made successfully.The change of expression of C1q protein and mRNA were observed with immunohistochemical staining and RT-PCR 15 days later. Results Immunohistochemically, the cell number of C1q positive staining in hippocampus of the AD model group（67.89±9.22） increased obviously as compared with the control group（36.27±5.09）（EM>P/EM>0.01）, and their average optical density（0.2918±0.0347） increased obviously as compared with the control group（0.1980±0.0183）（EM>P/EM>0.01）, the average optical density of C1q positive Related Articles | Metrics

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Effects of dihydrotestosterone on synaptic plasticity and the expression of N-methyl-D-aspartate receptor 1 in CA1 region of hippocampus in SAMP8 mouse 2010, 41 (6):  809-813.  doi: 10.3969/j.issn.0529-1356.2010.06.006 Abstract ( )   Objective To explore the effects of dihydrostestosterone(DHT) on synaptic plasticity and the expression of N-methyl-D-aspartate receptor 1（NMDAR1） in CA1 region of hippocampus in senescence accelerated mouse prone strain 8(SAMP8). Methods Twenty-one 6-month-old male SAMP8 were randomly divided into sham-operation control group, castrated group and DHT replacement therapy after castration group（7 in each group）. The dose of DHT was 1mg/(kgI>&#/I>8226;d) . Twentyone days later, the apical dendritic thorns density in hippocampal CA1 region was observed with Golgi staining method. Immunohistochemical method and computer pathological image analysis system were used to determine the expression of synaptophysin and N-methyl-D-aspartate 1(NMDAR1) in hippocampal CA1 region. Results 1. In the Golgi staining, the number of the apical dendritic thorns density of hippocampal CA1 region of castrated group decreased. However, DHT replacement therapy could significantly increase the apical dendritic thorns density (EM>P/EM><0.05). 2. The expressions of synaptophysin and NMDAR1 in the hippocampal CA1 region of castrated group decreased markedly. The average absorbance (AEM>A/EM>) values were sharply lower than those of other groups (EM>P/EM><0.05). DHT replacement therapy could obviously increase the expression of synaptophysin and NMDAR1 in the hippocampal CA1 region. Conclusion DHT replacement therapy can increase the density of dendritic thorns and modulate synaptic plasticity of hippocampal CA1 region. DHT may potentially affect hippocampal synaptic Related Articles | Metrics

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Motor neuron differentiation from neonatal mouse telencephalon neural stem cells 2010, 41 (6):  814-817.  doi: 10.3969/j.issn.0529-1356.2010.06.007 Abstract ( )   Objective To explore the possibility of motor neuron differentiation from neonatal mouse telencephalon neural stem cells and the novel inducing factors. Methods Neural stem cells were isolated from neonatal mouse telencephalon using floating culture methods. Three differentiation groups were divided according to different inducing factors. Group 1 containing growth medium+5% fetal bovine serum(FBS) served as control. Group 2 containing growth medium+5% FBS+ retinoid acid(RA)+Shh+dbcAMP was inducing factor group. Medium collected from skeletal muscle cultures was used for motor neuron induction in Group 3. Double immunofluoesence staining of microtubule associated protein 2(MAP2) and homeo box(HB9) were used to detect the differentiation of motor neuron. Twelve samples were selected randomly for cell counting. Results Motor neurons with MAP2 and HB9 coexpression were found in differentiation cultures. The ratio of motor neuron differentiation was 1% in Group 1, 4.7% in Group 2 and 2.9% in Group 3. The difference between Group 2 or Group 3 and Group 1 was of statistic significance. Conclusion Neonatal mouse telencephalon neural stem cells can differentiate into motor neurons. Skeletal muscle cells might secret motor neuron inducing factors. Related Articles | Metrics

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Age-related changes of gliosis in the medulla of primary somatosensory area of cat 2010, 41 (6):  818-822.  doi: 10.3969/j.issn.0529-1356.2010.06.008 Abstract ( )   Objective Agerelated morphological changes of glial cells, S100and glial fibrillary acid protein (GFAP)-immunoreactive cells in the medulla of primary somatosensory area (SI) were comparatively investigated between 4 young cats and 4 old cats, the causation and the effect for these changes were also discussed. Methods Modified Holzer staining was applied to show the total glial cells and the adult animal. Golgi method was used to display the cells’ morphology; Immunohistochemical method was employed to exhibit S100-immunoreactive (S100-IR) and GFAP-immunoreactive (GFAPIR) cells. Under microscope, the cell morphology was comparatively observed; The numbers of the glial cells, S100-IR and GFAP-IR cells were counted; the average optical density and the area ratio of the immunoreactive cells were calculated with the software of BI-2000. Results The density of the glial cells, S100-IR and GFAP-IR cells showed a significant increase in the medulla SI of old cats (EM>P/EM><0.01), accompanied with a notable increased ratio of S100-IR and GFAP-IR cells to the total glial cells. The average absorbance and the area ratio of the positive cells showed a remarkable increase in the old cats than those in the young ones (EM>P/EM><0.01). In addition，the GFAP-IR astrocytes exhibited larger soma and denser processes in the old animals. Conclusion The medulla of SI exists reactive gliosis during senescence, especially a more sensitivity of the astrocytes, which indicates that an increased glial activity might exert a protective function for aging nervous fibres and m Related Articles | Metrics

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Olfactory ensheathing cells transplanation reduced amyloid burden in amyloid precursor protein transgenic mouse model 2010, 41 (6):  823-826.  doi: 10.3969/j.issn.0529-1356.2010.05.009 Abstract ( )   Objective To observe the survival and migration of olfactory ensheathing cells (OECs) after transplanted into amyloid precursor protein (APP) transgenic mice and investigate its therapeutical effect on Alzheimer disease. Methods OECs from olfactory ensheathing bulb of enhanced green fluorescent protein(EGFP) transgenic mice were isolated, then transplanted into an APP transgenic mice. Stereotaxic coordinates and anterior-posterior were appilicated for microinjection. Confocal microscope was used to observe the survival and migration of the cells, immunohistochemistry and Western blotting were carried out to survey amyloid burden, 4 mice in each group. Results The results showed that OECs survived well and migrated away from injection site after transplanted into APP transgenic mice, the vascular phenomena of OECs was also observed, and the therapy reduced amyloid burden of Alzheimers disease model.Quantitative test showed that APP reduced significantly（EM>P/EM><0.05）. Conclusion Olfactory ensheathing cells do some help to reduce the amyloid burden in the brain of APP transgenic mice. Related Articles | Metrics

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Biocompatibility of silk fibroin with dog bone marrow stromal cells EM>in vitro/EM> 2010, 41 (6):  827-831.  doi: 10.3969/j.issn.0529-1356.2010.05.010 Abstract ( )   Objective To investigate the biocompatibility of dog bone marrow stromal cells (BMSCs) in vitro with silk fibroin material and to explore the possibility of using silk fibroin as a novel material in the fabrication of tissue-engineered nerve scaffolds combined with the introduction of BMSCs. Methods Dog bone marrow stromal cells were isolated from other cells by adherence to plastic. The dog BMSCs were then cultured on the substrate silk fibroin fibers and the cell attachment and growth observed using immunofluorescence light microscopy and scanning electron microscopy. The dog BMSCs were also cultured in the silk fibroin extract fluid. The cell ultrastructure was observed by transmission electron microscopy. MTT test was used to detect cell viability in different medium after 12, 24, 48 and 72 hours or 7 days in culture (EM>n/EM>=12 for each group). Flow cytometry was employed to detect BMSCs cell cycle and phenotypes (EM>n/EM>=3). Results BMSCs were tightly attached to the silk fibroin fibers and grew along the silk fibroin as demonstrated by immunofluorescence light microscopy and scanning electron microscopy, some of which exhibited a spherical, oval or spindle shape. The results of transmission electron microscopy, MTT test and flow cytometry analysis showed that there were no significant morphological, cell viability, proliferation and phenotypes differences between BMSCs cultured in the silk fibroin extract fluid and those in plain IMDM medium. Conclusion These data indicate that silk fibroin has good biocompatibility with BMSCs and does not exert any significant cytotoxic effects on their phenotype, thus it is a potential scaffold material combined with the introduction of BMSCs in the fabrication of tissue-engineered nerve. Related Articles | Metrics

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Gemcitabine (Gemzer) up-regulates the expression of gene EM>Tusc2/EM> and EM>Lats/EM> 2 in breast cancer mice 2010, 41 (6):  832-836.  doi: 10.3969/j.issn.0529-1356.2010.06.011 Abstract ( )   Objective To explore gemcitabine’s effect on tumor suppressor candidate 2(EM>Tusc/EM>2), large tumor suppressor2(EM>Lats/EM>2) gene expression in breast cancer of mice, both in mRNA and protein level. Methods Mice models of breast cancer were established and the samples were selected at 0 day, 5 days and 10 days after gemcitabine hydrochloride treatments as well as control. RT-PCR and Western blotting were used to detect the variation of the EM>Tusc /EM>2 and EM>Lats/EM>2 gene expression both for blank control, negative control and test groups. Results The results were similar both in RT-PCR and Western blotting. Compared with blank control, relative amounts of EM>Tusc/EM> 2 and EM>Lats/EM>2 RNA in test group increased around 4-5 and 8 times 10 days after treated with gemcitabine in breast cancer mice respectively (P<0.05). However, no differences were showed with Tusc2 and EM>Lats/EM>2 RNA level 5 days after treated with gemcitabine in breast cancer mice. As to protein expression level, TUSC2 and LATS2 protein relative amounts were raised around 5 and 10 times (P<0.05) compared with blank control. Significant statistical difference was fou Related Articles | Metrics

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Effects of periostin expression on the invasiveness of nasopharyngeal carcinoma cells EM>in vitro/EM> 2010, 41 (6):  837-841.  doi: 10.3969/j.issn.0529-1356.2010.06.012 Abstract ( )   Objective To investigate the effects and mechanism of periostin on their invasiveness of nasopharyngeal carcinoma cells EM>in vitro/EM>. Methods Western blotting was employed to detect the expression levels of periostin in the stroma of nasopharyngeal carcinoma (NPC) and normal nasopharyngeal mucosa (NNM), nasopharyngeal carcinoma cells 6-10B (low metastatic potential cell line ) and 5-8F (high metastatic potential cell line), NIH 3T3 fibroblasts. Eukaryotic expression plasmids of periostin were constructed and transfected into NIH 3T3 fibroblasts to obtain transient transfection cells, the expression of periostin was detected by Western blotting in NIH 3T3 cells before and after transfection. The biological effects were observed, including in vitro invasion by Boyden charmber assay, matrix metalloproteinase(MMPs) activation by gelatin zymogram. Results The expression levels of periostin protein varied in the two different metastatic-potential nasopharyngeal carcinoma cell lines and NIH 3T3 fibroblasts. Periostin was not expressed in 6-10B and NIH 3T3 cells, and 5-8F cells showed weak expression in comparison to the stroma of nasopharyngeal carcinoma. Boyden chamber assay indicated that the invasive capacity of 6-10B was significantly promoted by co-culture with Periostin-overexpression NIH 3T3 cells, and further study by gelatin zymogram assay indicated that MMPs in supernatant of periostin-overexpression NIH 3T3 cells co-culture with 6-10B were more active than negative controls and blank controls. Conclusion Periostin overexpression may promote the invasive capacity of nasopharyngeal carcinoma cells EM>in vitro/EM>. The action may exert through regulating the activity of MMPs. The study will provide a new idea for finding stromal targets in tumor therapy，and periostin is expected to be a new target Related Articles | Metrics

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Influence of progesterone on the number of the mouse uterine macrophages and the expression of its membrane receptors CD14 and CD204 2010, 41 (6):  842-846.  doi: 10.3969/j.issn.0529-1356.2010.06.013 Abstract ( )   Objective To study the influence of progesterone on the number of the mouse uterine macrophages and the expression of its membrane receptors CD14 and CD204, and to explore the role of pro-gesterone and macrophage in uterus regional immunity. Methods 1. Twenty-five non-pregnant and healthy mice were randomly and evenly divided into blank control group, negative control group and experimental group which divided into low, medium and high progesterone concentration subgroups in advance and every group had 5 mice. The progesterone at the doses of 0.4mg, 2.0mg, and 4.0mg were intracutaneously injected into the mice of the experimental group, respectively, while the same doses of placebo were injected into the negative control group in the same condition and both were injected continuously for 5 days; the blank control group was stayed intact. 2.Gestational day 4 to 5 mice, the 15, were also divided into blank control, negative control and experiment groups randomly, 5 mice in each group. The progesterone at the dose of 4mg was intracutaneously injected into the experimental mice in the morning, and as well the same doses of placebo were injected into the negative control group mice; the blank control group was left intact. All of the above mice uteri were collected 6 hours later after the final injection. Non-specific esterase(α-NAE) staining and immunohistochemistry methods were used to identify the varieties of α-NAESUP>+/SUP> ,CD14SUP>+ /SUP>,CD204SUP>+/SUP>macrophages in the uterus. Results Compared with the negative control group, after injecting progesterone, the amount of the uterine macrophages with positive CD14 receptor immunohistochemical staining were significantly decreased with dose increasing in the non gestational group. For the gestational day 4 to day 5 mice, the amounts of the uterine macrophages with the positive CD14 and CD204 receptors immunohistochemical staining were significantly increased(EM>P/EM><0,01). Conclusion Progeterone can affect the regional immunity of uterus by adjusting the quantity of macrophages or its distribution or the expression of its membrane receptor. And the macrophages from alternative activation pathway might have a contribution to the maintenance of the maternofetal immune tolerance. Related Articles | Metrics

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P>Specific down-regulation of glucose-regulated protein 78 decreases the invasion and metastasis capability of salivary adenoid cystic carcinoma/P> 2010, 41 (6):  847-851.  doi: 10.3969/j.issn.0529-1356.2010.06.014 Abstract ( )   Objective To explore the effect of specifically down-regulating glucose-regulated protein 78 on invasion and metastasis capability of adenoid cystic carcinoma. Methods Applying small interfering RNA (siRNA) technology to specifically downregulate the expression of GRP78 and analyzing the difference between ACC-2 and si-GRP78 treated ACC-2 by wound healing, Transwell assay, MTT and cytoskeleton staining. Then we detected the effect of down-regulating GRP78 in ACC-2 on matrix metalloproteinases(MMP)-2,MMP-9,tissue inhibitors of metalloproteinase(TIMP)1 and TIMP-2 by Western blotting technology to compare the ACC-2, control siRNA treated ACC-2 and siGRP78 treated ACC-2. Results Wound healing assay revealed down-regulating GRP78 in ACC-2 could inhibit the wound healing. From Transwell assay, we found that the penetrated cells number decreased significantly by down-regulating GRP78. Cytoskeleton staining showed the fibers of si-GRP78 treated ACC-2 were thickening and distinct obviously, and the fibers of ACC-2 and control siRNA treated ACC-2 were thin and blurred. MTT assay showed that down-regulating GRP78 could inhibit 1/3 reproductive ability of ACC-2. Western blotting manifested that down-regulating the expression of GRP78 in ACC-2 decreased the expression of MMP-2, MMP-9 and increased the expression of TIMP-1, TIMP-2. Conclusion Specific down-regulation of GRP78 could inhibit ACC invasi Related Articles | Metrics

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Comparison between nicotinamide and rat pancreatic extract for inducing mouse mesenchymal stem cells to differentiate into insulin producing cells 2010, 41 (6):  852-856.  doi: 10.3969/j.issn.0529-1356.2010.06.015 Abstract ( )   Objective By comparing rat pancreatic extract(RPE) with nicotinamide, we explored a more effective method for inducing mouse bone marrow mesenchymal stem cells(BMSCs) into insulin producing cells. Methods RPE and nicotinamide were used to trans-differentiate mouse BMSCs into insulin producing cells. After trans-differentiation, the two methods induced insulin producing cells(IPCs) were detected by immunocytochemistry to compare the insulin expression in cytoplasm and detected trans-differentiation extent by diphenylthiocarbazone(DTZ) staining. Insulin releasing curve of IPCs exposed to 25mmol/L glucose and C peptide expression were detected by Radioimmunoassay(RIA). Four pancreatic islets associated mRNA (Ins-1,Ins-2,Glut-2 and GK) expression by RT-PCR technology. Then we analyzed and compared trans-differentiation effect between the two methods. Result From immunocytochemistry of insulin, we found that insulin producing cells trans-differentiated by both methods and all could secrete insulin and no significant staining extent difference was observed. And also there was no difference between them in DTZ stain and morphology pictures. However RIA showed the evident difference between them. The insulin releasing curve by the IPCs exposed to 25mmol/L glucose showed that nicotinamide induced IPCs were not like RPE induced cells which were more similar to mouse pancreatic islets. Moreover, the nicotinamide induced BMSCs could not express C peptide, and the RPE induced BMSCs expressed C peptide. RT-PCR showed nicotinamide and RPE induced BMSCs could all express the islets related mRNA (Ins-1, Ins-2, Glut-2, GK). Conclusion As compared with Nicotinamide, RPE seems to be a more effective inducer to trans-differentiate BMSCs into IPCs. Related Articles | Metrics

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Expression of a FGF signalling inducible gene EM>Fin/EM>15 during early mouse tooth development 2010, 41 (6):  857-861.  doi: 10.3969/j.issn.0529-1356.2010.06.016 Abstract ( )   Objective To investigate the function of EM>Fin/EM>15 in mouse tooth development. Methods The expression of EM>Fin/EM> in mouse tooth germ was detected by RT-PCR. The expression of a fibroblast growth factor(FGF), inducible gene Fin15 in the early mouse dveloping tooth was detected by EM>in situ/EM> hybridization and EM>Fin/EM>15 expression in mouse mandible arch which was disrupted FGF singaling was examined by whole mount EM>in situ/EM> hybridization. Results EM>Fin/EM>15 was expressed in the both dental epithelium and mesenchyme from the initiation stage to the morphogenesis stage. Disruption of FGF signaling in mandible arch in organ culture abolished EM>Fin/EM>15 expression in the mandible, including tooth forming regions. Conclusion EM>Fin/EM>15 was a downstream target of FGF signaling pathway in developing tooth, implicating a role for EM>Fin/EM>15 in mediating function of FGF signaling in mammalian tooth Related Articles | Metrics

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Hair follicle growth cycle and the expression of β-catenin in mice 2010, 41 (6):  862-866.  doi: 10.3969/j.issn.0529-1356.2010.06.017 Abstract ( )   Objective To observe the histological characteristics of murine hair follicle in distinct hair follicle cycle stage, and to research the function of β-catenin in murine hair follicle cycle by investigating the expression and localization of β-catenin in murine distinct hair follicle cycle stage. Methods HE staining was used to observe the morphology of the murine hair follicle in each stage（EM>n/EM>=90）, Western blotting and immunohistochemistry were used to determine the expression level and localization of β-catenin protein in distinct hair follicle cycle stage. Results For each stage, stage-specific characteristics were provided in order to classify murine hair follicle cycle. The distribution of β-catenin in murine skin was demonstrated. The positive reaction of β-catenin was detected in epidermis, sebaceous glands, root sheath and dermal papilla. Conclusion The growth of murine hair follicle shows cyclical variation. The expression of β-catenin is significantly different in distinct hair follicle cycle stage suggesting that telogen-to-anagen transition depends on th Related Articles | Metrics

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P>Effects of compound panax notoginseng on the expressions of androgen receptor and c-kit in testicular tissues in formaldehyde rats exposure/P> 2010, 41 (6):  867-871.  doi: 10.3969/j.issn.0529-1356.2010.06.018 Abstract ( )   Objective To observe the morphological variation of seminiferous tubules and the expressions of the androgen receptor(AR) and tyrosine kinase receptor c-kit in the testicular tissue in formaldehyde exposure SD rats after being treated with compound panax notoginseng(CPN). Methods Eighteen male SD rats were randomly divided into three groups：control，formaldehyde and CPN group．Formaldehyde and CPN group were stayed in the chambers filled with formaldehyde vapor（69.5±8.3ppm）for 8 weeks（8hours/day）. From the fifth week, the rats in CPN group were treated with CPN(1ml/100g) everyday for 4 weeks. At the end of the experiment, the whole body and testes of all rats were weighed. The seminiferous tubules and the expressions of androgen receptor and c-kit in testicular tissues observed with HE staining and the immunohistochemical method,respectively. Results The weights of bodies and testes in the formaldehyde group decreased significantly in comparison with the control group.After treated with CPN for 4 weeks, both weights significantly increased compared to the formaldehyde group(EM>P/EM><0.05). The atrophy of the convoluted seminiferous tubules，disorderly arrangement, enlarged interval of epithelial cells of the tubules and the remarkable loss of the sperms were observed in the formaldehyde group using HE staining. The morphology of seminiferous tubules gradually recovered, and lots of spermatids appeared in the seminiferous tubules in CPN group.The expressions of androgen receptor and c-kit in testicular tissues decreased significantly in formaldehyde group, but increased remarkably almost to the normal level in CPN group(EM>P/EM><0.05). Conclusion Formaldehyde exposure for a long time causes the loss of the weights of bodies and testes and the declines of the expressions of androg Related Articles | Metrics

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Effect of oxidative stress on diabetic erectile dysfunction in rats 2010, 41 (6):  872-875.  doi: 10.3969/j.issn.0529-1356.2010.06.019 Abstract ( )   Objective To investigate the effect of oxidative stress on diabetic erectile dysfunction( ED). Methods Diabetes mellitus(DM) was induced by streptozotocin (STZ) injection. Eight weeks and twelve weeks later, penis erectile function was detected by injecting apomorphine. The concentrations of malondialdehyde ( MDA) and total anti-oxygen capability (T-AOC) in penis were measured. Western blotting was used to measure nuclear factor E2related factor 2（Nrf2） in both control and diabetic rat penis tissues in present study. Results Compared with control group, the erection times of DM rats decreased significantly. The mean level of MDA in penis of DM rats were higher than that of control group（EM>P/EM>﹤0.01）, and the mean level of T-OC in DM rats was lower than that of control（EM>P/EM>﹤0.05）. Nrf2 protein expression decr Related Articles | Metrics

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Effect of human foreskin fibroblast as a feeder layer on growth of human embryonic germ cells 2010, 41 (6):  876-879.  doi: 10.3969/j.issn.0529-1356.2010.06.020 Abstract ( )   Objective To culture human embryonic germ cells (EGCs) on feeder layer of human foreskin fibroblast (HFF), and observe the growth of EGCs and identify EGCs. Methods To separate and culture HFF of children aged 4-5, the fibroblasts from third generation to 30th were used in the experiment. The fibroblast growth factor(FGF) concentration in HFF culture medium was detected by ELISA-double antibody sandwich assay. The primordial germ cells were isolated from 5-11 weeks human embryos. The EGCs were cultured in medium without exogenous growth factors, only supported by HFF as feeder layers．The activity of alkaline phosphatase, specific antigen of the embryo SSEA-1 and SSEA-4 and Oct-4 were respectively detected by cytochemistry, immunocytochemistry and RT-PCR. Results The fibroblasts, with an anticipated life span were of 60 generations, were used in our experiments after 3-30 generations. The concentrations of FGF in HFF culture medium were ranged from (172.09±2.66) pg/L to (245.25±1.6) pg/L. Human EGCs cultured in the HFF feeder layer can form typical embryo germocyte colonies. After having been cultured for P3 generations continuously, the activity of alkaline phosphatase of EGCs, undifferentiation marker SSEA- and SSEA- of EGCs colonies and the expression of transcription factor Oct- showed positive. Conclusion HFF can be a feeder layer to support the growth of human EGCs and maintain its undifferenti Related Articles | Metrics

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Effect of heroin withdrawl and relapse on expression of 5-HT,gastrin,somatostatin immunoreactive cells in duodenum of rats 2010, 41 (6):  880-884.  doi: 10.3969/j.issn.0529-1356.2010.06.021 Abstract ( )   Objective To study the changes of expression of 5-hydroxy tryptamine(5-HT), gastrin(Gas) and somatostatin(SS) in rat duodenum during heroin abstinence and relapse period for investigating the effects of the possible functions of 5-HT, Gas, SS in duodenum during heroin abstinence and relapse. Methods Thirty-five SD adult male rats were randomly divided into three groups: normal control group (NCG), saline control group (SCG) and experiment group (EG). Experiment group was divided into: heroin abstinence group(HAG), methadone detoxication group(MDG) and heroin relapse group(HRG). Duodenal tissues were obtained from three groups(NCG,SCG,EG) separately. Immunohistochemical SABC method was adopted, cell counting and image analysis were used in the research. Results Compared with NCG and SCG, the number of 5-HT,Gas,SS immunoreactive cells increased (EM>P/EM><0.05),the results of image analysis showed that the mean grey degree of 5-HT, Gas, SS immunoreactive cells decreased obviously in HAG and HRG (EM>P/EM><0.05). Compared with NCG and SCG, there were no statistical significances on the 5-HT,Gas,SS immunoreactive cells counting (EM>P/EM>﹥0.05) in MDG. Conclusion In HAG and HRG, 5-HT, Gas, SSimmunoreaction intensity and the number of cells increased obviously in rats duodenum, suggesting that 5-HT, Gas, SS participate in the process of adjusting during HAG and HRG. After detoxication of methadone, the expression of 5-HT, Gas, SS was close to normal, suggesting that the changes of expression of 5-HT, Gas, SS were reversible. Related Articles | Metrics

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Atelocollagen collagen scaffold is facilitated to cultured three-dimensional myocardium of neonatal rat EM>in vitro/EM> 2010, 41 (6):  885-890.  doi: 10.3969/j.issn.0529-1356.2010.06.022 Abstract ( )   Objective To culture three-dimensional contractible myocardial cell block of neonatal rats in atelocollagen scaffold and to further establish the foundation for heart tissue engineering. Methods After being cultured and purified, the neonatal rats’ cadiocytes were planted into atelocollagen scaffold for three-dimensional (3D) cultures. The growth of cadiocytes in atelocollagen scaffold was dynamically observed by inverted microscopy. Myocardial cell blocks in different periods were separately tested by HE staining, immunohistochemistry and immunofluorescence of myoglobulin, and observed under transmission electron microscope. Results After being planted, cadiocytes began to grow on atelocollagen scaffold in 6 hours, appeared mutual integration of myocytes and formed a complex with the scaffold to pulsate together on the 2nd day. And then, mutual integration among cadiocytes binding became more closely than before on the 6th day, and cadiocytes emerged into reticulation in the scaffold meshes on the 10th day. On day 20 of the culture,cadiocytes filled most of atelocollagen scaffold meshes and formed a tight block of myocardial tissue. The cadiocytes were always kept well the contractility and autorhythmicity in the whole course of culture. The growth cells in scaffold meshes were mainly cardiomyocyte, identified by morphological methods, with very few fiberlike cell. Conclusion Cadiocyte cultured in atelocollagen scaffold could develop the ideal and three-dimensional myocardial cell block with systolic function. Related Articles | Metrics

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Change of Rho associated kinase-1 and epithelial-mesenchymal transition in mouse pulmonary fibrosis 2010, 41 (6):  891-896.  doi: 10.3969/j.issn.0529-1356.2010.06.023 Abstract ( )   Objective To investigate the developing expression of Rho associated kinase-1(Rock-1) and its relation to epithelial-mesenchymal transition in bleomycin-induced pulmonary fibrosis in mice. Methods Sixty mice were randomly divided into three groups, control group(EM>n/EM>=20)(normal saline),experimental group (EM>n/EM>=20)( blemycin-injected 5mg/kg,intratracheal injection) and treated group (EM>n/EM>=20)(dexamethason-injected 5mg/kg every day, intraperitoneal injection). On day 3,7,14 and 28 after treatment, lung tissue was stained with HE method.Lung hydroxyproline (HYP) content was evaluated. Immunohistochemistry was used to detect the expression of alpha-smooth muscle actin (α-SMA) and E-cadherin(E-cad). Western blotting was used to detect the expression of Rock-1. Results In experimental group, dynamic changes from alveolitis to pulmonary fibrosis could be observed in the slices by pathological method. Collagen deposition increased evidently on day 3, and peaked on day 28. Lung hydroxyproline (HYP) content increased gradually. Compared to those in the control group, the protein level of α-SMA increased significantly in the model group and treated group, while the protein level of E-cadherin decreased significantly. The expressions of Rock-1 protein in lung tissues in experimental group were higher than those in the control group(EM>P/EM>< 0.05),which increased from the third day, and peaked on the 14th day, then decreased on the 28th day. While the protein level of Rock-1 in treated group decreased significantly. Conclusion The expression of Rock-1 is positively correlated with not only the degree of pulmonary fibrosis but also the level of α-SMA and E-Cad, which probably suggests that Rock-1 is a promoter to pulmonary fibrosis via the induction of epithelial-mesenchymal Related Articles | Metrics

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Three-dimensional reconstruction of the mouse renal proximal tubule 2010, 41 (6):  897-900.  doi: 10.3969/j.issn.0529-1356.2010.06.024 Abstract ( )   Objective To characterize the three-dimensional (3D) course of the mouse proximal tubules. Methods Three C57/BL/6J mice were fixed by perfusion and embedded in Epon 812. Tissue blocks were cut perpendicular to the longitudinal axis of the kidney, then a total of 1 200, 2.5-μm-thick consecutive sections were obtained from the surface to the outer stripe of the outer medulla. After image recordings and alignment, proximal tubules from 58 nephrons were traced and 3D-reconstructed with a series of computer programs written in C language. Results In cortical labyrinth, all of the proximal tubules left their glomeruli ascending toward the renal surface for a distance of 100-1 400μm, then returned and made coils around their own glomeruli and occupied separate domains which did not intermingle with the neighboring nephron tubules. The proximal tubules originating from the superficial nephrons and midcortical nephrons formed tight clusters and occupied smaller volume than the proximal tubules originating from the juxtamedullary nephrons. In medullary rays, the pars recta were arranged in a specific pattern: proximal tubules originating in the superficial cortex were arranged in the center, whereas proximal tubules originating deeper in the cortex became layered more peripherally, and in the deepest juxtamedullary nephrons, it was impossible to define a pars recta since the “straight” part did not run straightly in the outer stripe of the outer medulla. All the proximal tubules terminated at the boundary between the outer stripe of the outer medulla and the inner stripe of the outer medulla and transitioned to descending thin limb of the Henle’s loop. Conclusion In the cortical labyrinth and medullary rays, the courses of the initial, convoluted and straight proximal tubules run in different regions and in different biological environment, which supply implications for evaluting segment distributio Related Articles | Metrics

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Histological observation and three-dimensional reconstruction of human modiolus 2010, 41 (6):  901-904.  doi: 10.3969/j.issn.0529-1356.2010.06.025 Abstract ( )   Objective To further research the histological composition and characteristics of modiolus by histological observation and three-dimensional recons-truction. Methods Four tissue samples lateral to the angle oris were paraffin imbedded and sectioned, stained with Verhoeff iodine-hematoxylin, observed under the microscope. One tissue sample image among them was put into Winsurf software on personal computer, and then was three-dimensional reconstructed. Results Modiolus in histology was found in orbicularis oris，zygomaticus major muscles，levator anguli oris，depressor anguli oris，platysma pars modiolus and risorius converged in the superficial layer of buccinator lateral to the angle of the mouth. Collagens were thick conspicuously where the muscles converged, and elastic fibers intermixed in it.The thickness of modiolus was 2.52mm, and the apex moduli was ellipse like.Measurements of three-dimensional reconstruction model were carried out. The results were as follows: distance between center point of apex moduli and angle oris was 11.96mm; volume of modiolus was 84.10mmSUP>3/SUP>; perimeter of apex moduli was 27.13mm, area was 18.41mmSUP>2/SUP>；perimeter of base moduli was 33.39mm, area was 28.86mmSUP>2/SUP>. Conclusion Relative to the familiar definition of modiolus in gross anotomy, definition of modiolus in histology which observes under microscope is suggested, also its three-dimensional model is reconstructed, then Related Articles | Metrics

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Correlation among scan parameter, image quality and radiation dose of cadaver wrist multislice spiral computed tomography 2010, 41 (6):  905-908.  doi: 10.3969/j.issn.0529-1356.2010.06.026 Abstract ( )   Objective To study correlations among scan parameter, image quality and radiation dose of cadaver wrist multislice spiral computed tomography(MSCT). Methods Four cadaver wrists labeled as 1-4 respectively underwent 16-slice CT (Siemens Somatom Sensation16) scan. Totally 11 groups of MSCT scan were performed for each specimen.Scan parameters were different. An additional scan was performed for the third specimen with a high voltage(140kV) and a high current(300mAs).Thin-slice coronal images were reformatted for all the scans.Each reformatted coronal image was evaluated. Correlations among scan parameter, image quality and radiation dose were analyzed. Results Image quality of each wrist with different scan parameters did not show evident difference. Radiation dose increased linearly with current and increased curvedly with voltage. Radiation dose did not change obviously when pitch was changed. Conclusion Relations are different between radiation dose and different scan parameters. Correlations among scan parameter, image quality and radiation dose are instructive in practice. Related Articles | Metrics

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Effects of aloe-emodin on proliferation and migration of human gastric cancer cell line BGC-823 2010, 41 (6):  909-911.  doi: 10.3969/j.issn.0529-1356.2010.06.027 Abstract ( )   Objective To investigate the effect of aloe-emodin on proliferation and migration of human gastric cancer cell line BGC-823. Methods The effects of aloe-emodin on cell proliferation,cell cycle stage and migration ability of BGC-823 cells were observed.The proliferation level was measured by MTT method, cell cycle stage was detected with flow cytometry and migration ability was tested by wound healing assay. Results MTT assay showed the inhibitory effect of aloe-emodin on proliferation of BGC-823 cells was dose-dependent; Treatment of BGC-823 cells with aloe-emodin resulted in cell cycle arrest at GSUB>0/SUB>/GSUB>1/SUB> phase by flow cytometry analysis; The migration velocity of BGC-823 cells was obviously inhibited as shown in wound healing assay. Conclusion The inhibitory effect of aloe-emodin on proliferation and migration of BGC-823 cells was dose-dependent, as well as the cell cycle was arrested at GSUB>0/SUB>/GSUB>1/SUB> phase, which offers the clue for further understanding the inhibition mechanism of aloe-emodin on human gastric cancer cell line BGC-823. Related Articles | Metrics

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Role of Rho kinase in morphology and migration of cultured Schwann cells based on time-lapse imaging 2010, 41 (6):  912-915.  doi: 10.3969/j.issn.0529-1356.2010.06.028 Abstract ( )   Objective To investigate the effect of Rho kinase (ROCK) on the morphology and migration of cultured Schwann cells. Methods Primary culture Schwann cells were performed and the purity of Schwann cell was identified by immunofluorescence. Single-cell migration assay for Schwann cells was established. Based on this assay, a gradient of ROCK inhibitor Y27632 was produced in front of migrating Schwann cells. The behavior of morphology and motility in Schwann cells by Y-27632 were recorded based on time-lapse imaging. Results Immunofluorescence showed that cultured cells were of high purity of Schwann cell and single-cell migration assay was suitable for studying the morphology and motility of Schwann cells. Y-27632 dramatically promoted the more braches, thinner and longer processes, and inhibited Schwann cell migration. Conclusion ROCK can maintain the morphology and regulate the motility of Schwann cells. Related Articles | Metrics

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Effect of eucalyptus on embryonic development of rats 2010, 41 (6):  916-918.  doi: 10.3969/j.issn.0529-1356.2010.06.029 Abstract ( )   Objective To study the effects of eucalyptus on embryonic development of rats. Methods Totally 25 pregnant SD rats were randomly divided into 5 groups, each had 5 rats: three experiment groups ( high，medium，low doses of eucalyptus), negative control group ( peanut oil), positive control group (cyclophosphamide). Three experiment groups and negative control group were lavaged from gestation 10 to 14 days with eucalyptus in 100mg/(kgI>&#/I>8226;d), 200mg/(kgI>&#/I>8226;d), 300mg/(kgI>&#/I>8226;d) and peanut oil 2ml per animal per day respectively, while positive control group was ip with 12.5mg/(kgI>&#/I>8226;d) cyclophosphamide at gestation 13 days.All pregnant rats were sacrificed at the 19th day. The body, uterus, ovary and placenta weights of the pregnant rats were measured respectively. The numbers of absorbed, live and dead fetus were counted, respectively. The length for fetus body and tail as well as body weights were measured. Results After administration of eucalyptus (100mg/kg,200mg/kg,300mg/kg) for 5 days, the body, uterus, ovary and placenta weights of pregnant rats showed no significant difference compared with negative control group(EM>P/EM>>0.05).The numbers for absorbed, live and dead fetus also showed no significant difference compared with negative control group ( EM>P/EM>>0.05 ). All of the fetus body weights, body length and tail length were larger in the treated groups than those in the negative control group. But only in high doses of eucalyptus groups, the fetus weight of body showed difference compared with negative control group (EM>P /EM><0.05), others had no significant difference (EM>P/EM>> 0.05). The fetus body weight, tail length in the positive control group were lower than those in the negative control group (EM>P/EM> <0.05),but the fetus body length showed no significant difference compared with negative control group (EM>P /EM>>0.05). Conclusion Under the experimental condition, eucalyptus showed no embryonic developmental toxicity, where cyclophosphamide showed embryonic developmental toxicity. Related Articles | Metrics