Table of Content

2011, Volume 42 Issue 6
06 December 2011 Previous Issue    Next Issue
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神经生物学
细胞和分子生物学
肿瘤生物学
解剖学
组织学胚胎学发育生物学
人类学
生物工程学
技术方法


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神经生物学

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Fimbria fornix transected hippocampal extracts promote proliferation and differentiation of radial glia cells EM>in vitro/EM> 2011, 42 (6):  721-725.  doi: 10.3969/j.issn.0529-1356.2011.06.001 Abstract ( )   Objective To investigate the proliferation and differentiation effect of hippocampal niche on the radial glia cells EM>in vitro/EM> . Methods Successfully prepared normal hippocampal extracts and transected extracts after hippocampal fimbria-fornix transection. On the proliferation and differentiation stages, the normal and transected extracts were used to investigate their effects on the radial glia cells respectively. Results On the proliferation stage, the proportion of BrdU-positive cells in the transected group was obviously higher than that of the normal group and control group（EM>P/EM>0.05）; on the differentiation stage, the number of Tuj1-positive cells in the transected group was much more than that of the normal and control group（EM>P/EM>0.05）. Conclusion The fimbria-fornix transected hippocampal extracts can promote the proliferation and neurons differentiation of neonatal rats radial glia cells EM>in vitro/EM>. These data indicate Related Articles | Metrics

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erived neurotrophic factor overexpression promoting the differentiation of rat neural stem cells into neurons 2011, 42 (6):  726-730.  doi: 10.3969/j.issn.0529-1356.2011.06.002 Abstract ( )   Objective To construct eukaryotic expression vector of brain-derived neurotrophic factor (BDNF) and detect its effect of overexpression on differentiation of rat neural stem cells (NSCs) into neurons. Methods The RT-PCR was used to amplify rat BDNF gene from RNA of rat hippocampus. The BDNF gene was inserted into eukaryotic expression vector pEGFP-N1 to construct recombinant expression vector pEGFP-N1-BDNF. The recombinant vector was transfected into NSCs by Lipofectamine 2000.The expression of BDNF mRNA in NSCs was detected by RT-PCR. The differentiation of rat NSCs into neurons was detected by immunohistochemistry staining. Results The sequence of the cloned BDNF was confirmed to be correct by DNA sequencing. The NSCs transfected with pEGFP-N1-BDNF expressed BDNF efficiently. The pEGFP-N1-BDNF transfected NSCs differentiated into more neurons than the pEGFP-N1 transfected ones (EM>P/EM>0.01). Conclusion All these results indicate that BDNF overexpression significantly promotes the differentiation of rat NSCs Related Articles | Metrics

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Inhibition effects of transplantation of bone marrow stromal cells modified by GDNF gene on neural cell apoptosis in rats with intracerebral hemorrhage 2011, 42 (6):  731-736.  doi: 10.3969/j.issn.0529-1356.2011.06.003 Abstract ( )   Objective To explore the effects of bone marrow stromal cells (BMSCs) transplantation modified by glial cell line-derived neurotrophic factor (GDNF) gene on neuronal apoptosis and the expression of apoptosis-related genes in rats with intracerebral hemorrhage. Methods Forty-eight rats were used to establish the model of intracerebral hemorrhage by injecting with collagenase and heparin into caudate nucleus through stereotaxic apparatus.The rats were randomly divided into BMSCs group, GDNF/BMSCs group and saline group, and were stereotaxically grafed with BMSCs, GDNF/BMSCs and saline respectively at the 3rd day after operation. Each group was subdivided into two subgroups according to different refeeding time (1 week, 2 weeks). Neurological function and area of brain injury were assessed by neurological deficits score and HE staining respectively. Expression of GDNF mRNA was observed by RT-PCR. The number of neuronal apoptosis and the expressions of Bax and Bcl-xl in the margin of the hemorrhagic focus were observed by TUNEL and immunohistochemistry. Results Significant recovery of neurological function and increase of GDNF mRNA expression were found in the GDNF/BMSCs group compared with BMSCs and saline group. Both rate of brain injury area and the number of neuronal apoptosis in the GDNF/BMSCs group were significantly less than other groups. As compared with the BMSCs and saline group, the number of Bcl-xl positive cells increased in the GDNF/BMSCs group, while the number of Bax positive cells decreased. Conclusion The transplantation BMSCs modified by GDNF gene provides better neuroprotection than native BMSCs. The underlying mechanisms may be due to partly inhibited apoptosis, and the expression of up-regulated Bcl-xl and down-regulated Bax protein. Related Articles | Metrics

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Immunohistochemical localization of DARPP-32, a dopamine and cyclic-AMP-regulated phosphoprotein, in the rat brain 2011, 42 (6):  737-740.  doi: 10.3969/j.issn.0529-1356.2011.06.004 Abstract ( )   Objective To study the distribution of DARPP-32 in the rat brain. BR>Methods Immunohistochemical staining techniques were applied to study the localization of DARPP-32 in neurons of the rat brain. Results The strong immunoreactivity for DARPP-32 was localized primarily in the basal ganglia and the anterior olfactory area, involving neuronal cell bodies or dendrites in the nucleus accumbens, caudate putamen, amygdaloid nucleus and fibers throughout the globus pallidus, ventral pallidum and pars reticulate of the substantia nigra. The moderate to weak immunoractivity for DARPP-32 was observed in red nucleus, septofimbrial nucleus, purkinje cells in cerebellum, medial habenula nucleus, cortex and hippocampus. DARPP-32 was present in the cytoplasm, cytomembrane and cell nucleus of labeled neuronal somata and dendrites. Conclusion DARPP-32 is widely distributed in the rat brain. It is primarily present in the neurons, which receive a dopamine input and contain dense of the D1 receptor. DARPP-32 may play an important role in dopaminoceptive neurons. BR> Related Articles | Metrics

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Effects of transient hypoxia on apoptosis and neurogenesis in the newborn rat brain 2011, 42 (6):  741-745.  doi: 10.3969/j.issn.0529-1356.2011.06.005 Abstract ( )   Objective To investigate effects of transient hypoxia on the expression level of neurogenic differentiation factor (NeuroD), apoptosis and neurogenesis in the rat brain.Methods The model was established as previously described by Grojean, newborn rats were transiently exposed to 100% N2. Ninety-six animals were randomly divided into normal control group and 20min hypoxia-treated group. Rats were sacrificed at day 13, 20 and 27 post-hypoxia, and the expression levels of NeuroD and BrdU in the brain were analyzed by immunofluorescence staining and immunohistochemistry. Alternatively, the rats were sacrificed at day 6 and 20 post-hypoxia, and apoptosis in the brain was analysed with TUNEL staining. Results Immunofluorescence/immunohistochemisty analysis showed that the expression of NeuroD and BrdU significantly increased at day 13 and 27 in the 20min hypoxia-treaded group compared to that in the control group (P0.05), with the highest expression level at day 20 (P0.01). TUNEL positive cells also significantly increased at day 6 in the 20min hypoxia-treaded group, while no difference was seen between the hypoxia-treated and the control group at day 20. Conclusion The transient neuronal loss induced by birth hypoxia may lead to cell proliferation, neuron differentiation and also may trigger neurogenesis, which may in turn participate in brain repai Related Articles | Metrics

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Expression of brain lipid binding protein in rat hippocampus dentate gyrus after traumatic brain injury 2011, 42 (6):  746-750.  doi: 10.3969/j.issn.0529-1356.2011.06.006 Abstract ( )   Objective To investigate the change of brain lipid binding protein(BLBP) expression in hippocampus dentate gyrus(DG) at different time points after traumatic brain injury (TBI). Methods Seventy-two rats were divided into the injured group, sham group and control group randomly, and then were subjected to a lateral fluid percussion injury (FPI). Western blotting was used to detect BLBP expression. Brains were sectioned for immunofluorescence staining of BLBP and Vimentin at the time points of 1, 3, 7, 14 days after TBI.Results The results of Western blotting showed that the BLBP expression was lower than that of control group at 1 day post injury（EM>P/EM><0.01） and reached the peak compared with the other groups at 7 days after injury（EM>P/EM><0.01）, then descended at 14 days compared with control group after injury（EM>P/EM><0.01）. The changes of BLBP and Vimentin double-label positive cells were consistent with the results of Western blotting. The BLBP and Vimentin double-label positive cells were found mainly at the subgranular zone of ipsilateral injured hippocampus DG, and most of them were radial glia like cells; BLBP and Vimentin double-labelled positive cells were found at the hilus of DG at 7days after injury, and most of them looked like reactive astrocytes. Conclusion The expression of BLBP in DG after TBI decreased firstly, then increased and reached peak at 7 days after injury, decreased dramatically again at last. The Related Articles | Metrics

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Transection of the fimbria fornix inducing neurogenesis in its proximal portion in adult rats 2011, 42 (6):  751-755.  doi: 10.3969/j.issn.0529-1356.2011.06.007 Abstract ( )   Objective To determine the neurogenesis in injured fimbria fornix (FiFx).Methods The left fimbria fornix of SD rat ( EM>n/EM> =7) were subjected to transection followed by intraperitoneal injection of 5-bromo-2-deoxy-uridine (BrdU). Coronal sections through hippocampus were stained with Nestin and BrdU antibodies 7 days after operation. The normal and injured FiFx were dissociated for neural stem cell specific culture 7 days post transection. Immunofluorescence staining was used to determine the properties of the neurospheres.Results Immunofluorescence revealed that Nestin-expressing cells were present in FiFx following lesion. Majority of Nestin positive cells were also positive for BrdU, a marker of DNA synthesis. Nestin positive proliferative cells were almost entirely absent from unlesioned tissue. Neurospheres cultured in vitro from lesioned FiFx displayed the characteristics of neural stem cells (NSCs) proliferation, expression of embryonic markers, and multipotential differentiation into neurons, astrocytes and oligodendrocytes. Conclusion These results demonstrate that transection of the tracts related with hippocampal function can stimulate NSCs proliferation in the projection pathways themselves. Ectopic neurogenesis in FiFx may provide an additional mechanism for repair of innervation following damage. Related Articles | Metrics

细胞和分子生物学

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Establishment of Tetracycline-induced gene expression system based on the lentiviral vector 2011, 42 (6):  756-760.  doi: 10.3969/j.issn.0529-1356.2011.06.008 Abstract ( )   Objective To establish Tetracycline-induced gene expression system for gene therapy based on third generation lentivirus system.Methods The mutant of the reverse Tet transactivator (rtTA and M2rtTA) was subcloned into a lentiviral vector with neo selection marker named as pELNS-rtTA-IRES-Neo and pELNS-M2rtTA-IRES-Neo. The Tet-responsive element (TETO and TREpitt) and green fluorescence proteint (GFP) were subcloned into a lentiviral vector with blasticidin selection marker named as plenti6-TETO-GFP and plenti6-TREpitt-GFP.293 cells were contransfect with pELNS-rtTA-IRES-Neo and plenti6-TETO-GFP, or with pELNS-M2rtTA-IRES-Neo and plenti6-TREpitt -GFP. Cells were treated by Dox to induce GFP expression. After 48hours, GFP expression in the co-transfected cells was observed under a fluorescent microscope.Results The first generation of Tetracycline-induced gene expression system named pELNS-rtTA-IRES-Neo and plenti6-TETO-GFP vectors were successfully set up. GFP expression in cotransfected cells induced with Dox was about 90% positive, while there was 30% positive GFP expression observed in no Dox inducing group. The second generation of Tetracycline-induced gene expression system named pELNS-M2rtTA-IRES-Neo and plenti6-TREpitt-GFP vectors were successfully set up. GFP expression in cotransfected cells induced with Dox was about 95% positive, while there was no positive GFP expression observed in no Dox inducing group.Conclusion Tetracycline-induced gene expression system based on lentivirus was successfully set up, which can induce gene expression effectively and tightly without obvious side-effects on cells by using the second Tetracycline-induced elements. Related Articles | Metrics

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Id2 translocation from nucleus to cytoplasm accelerating differentiation of skeletal muscle cells by regulating the expression of apoptosis inducing factor 2011, 42 (6):  761-765.  doi: 10.3969/j.issn.0529-1356.2011.06.009 Abstract ( )   P>Objective To explore the functional role of Id2 in skeletal muscle regeneration. Methods Id2 expression vectors were transferred into C2C12 cells. The transferred and un-transferred C2C12 skeletal muscle cells were exposed to 50μmol/L H2O2 and 2% horse serum for 12 hours without fetal bovine serum(FBS). Expression of Id2 gene in transferred and untransferred C2C12 cells was observed by RT-PCR. Expression of various myogenesis related proteins in the transferred and untransferred C2C12 cells were observed by Western blotting. Expression of Id2 and AIF proteins in the normal, fiber-damaged and denervated skeletal muscles were observed by immunofluorescence.Results Compared with un-transferred cells, the Id2 transferred cells exhibited higher differentiation. Immunofluorescence staining revealed that 50μmol/L H2O2 treatment increased the expression of nucleic Id2.Under the oxidative stress, Id2 repressed both MyoD repressors and myogenin activator. Two percents of the horse serum, which usually was used to induce myoblasts differentiation, caused most of Id2 proteins translocation from nucleus to cytoplasm. Translocation of Id2 protein from nucleus to cytoplasm inhibited the ROS-induced expression of mitochondrial apoptosis inducing factor(AIF). Immunofluorescence analysis implied that the denervated skeletal muscle showed more increased Id2 and AIF proteins in the nucleus.Conclusion Id2 translocation from nucleus to cytoplasm can accelerate differentiation of skeletal muscle cells. The functional role of Id2 during the skeletal muscle regene Related Articles | Metrics

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Smad3 gene promoting proliferation of GCs in rats 2011, 42 (6):  766-769.  doi: 10.3969/j.issn.0529-1356.2011.06.010 Abstract ( )   Objective To study the effects of Smad3 on the proliferation of the ovarian granulosa cells (GCS) in the rat.Methods Ovarian granulosa cells were cultured and treated in the flollowing conditions: 1. GCs were cultured without any treatment; 2. GCs were cultured with transfection reagent lipofectamine RNAiMAX; 3. GCs were transfected with transfection reagent lipofectamine RNAiMAX and Non-silencing SiRNA; 4. GCs were transfected with transfection reagent lipofectamine RNAiMAX and the SiRNA of Smad3.The expression and localization of PCNA and Cyclin D2 proteins in the four groups were detected by immuocytochemistry. Results The proteins of PCNA and Cyclin D2 were mainly located in the nuclei of GCs. The percentage of positive cells for PCNA was (25.80±1.70)% in the siRNA of Smad3 treated group, which decreased significantly compared to the control group(P <0.05),and the percentage of the stained cells for Cyclin D2 was (25.00±2.90)% in the siRNA of Smad3 treated group, which decreased significantly compared to the control group(P <0.05).Conclusion Smad3 gene could promote the proliferation of ov Related Articles | Metrics

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Decrease of serum protein concentration difference by solid-phase ligand libraries 2011, 42 (6):  770-774.  doi: 10.3969/j.issn.0529-1356.2011.06.011 Abstract ( )   Objective To determine the effects of solid-phase ligand libraries on enrichment of low-abundance serum protein and improve the low-abundance serum protein identification by mass spectrometer(MS).Methods Three pooled serum samples were treated with ProteoMiner and then subjected to one-dimensional (one-D) or two-dimensional (two-D) SDS-PAGE followed by comparative image analysis.The remarkable increased protein spots on two-D map were identified by high performance liquid cbromatogrphy(HPLC) coupled on-line to ESI/MS-MS.Results The one-D and two-D gel map revealed more low-abundance protein bands and protein spots, respectively. MS identification of significantly enriched protein spots on two-D gel improved low-abundance protein identification after library beads treatment, e.g. clusterin, vitronectin, C4b-binding protein alpha chain，complement C4-A. Conclusion The technique of solid-phase ligand libraries can enrich serum low-abundance proteins whilst decrease high-abundance protein concentration, which indicates that this method is useful for enrichment of low-abundance protein and id Related Articles | Metrics

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bFGF induced rat bone marrow mesenchymal stem cells differentiate into cardiomyocyte-like cells EM>in vitro/EM> 2011, 42 (6):  775-781.  doi: 10.3969/j.issn.0529-1356.2011.06.012 Abstract ( )   Objective To study the feasibility of basic fibroblast growth factor （bFGF）in inducing bone marrow mesenchymal stem cells(BMSCs) to differentiate into cardiomyocyte-like cells in vitro. Methods SD rat BMSCs were isolated and cultured from rat bone marrow, and then induced by bFGF. The cultured cells were observed by a phase-contrast microscope. The immunohistochemical technique and laser scanning confocal microscope (LSCM) were used for examination of the expression of desmin, α-actin and C-TnT. The ultrastructure of induced cells was observed by a transmission electron microscope. GATA4 andα-MHC expressions were detected by relative quantitative RT-PCR after 7, 21, 28 days of induction respectively. Results After primary cells were cultured for 48 hours，most of them became fusiform fibroblast-like shape with two or three processes and a few of them were flat in shape. The morphology of BMSCs induced by bFGF changed obviously. After being induced by bFGF for one week, the cells became larger and most of them became myocyte-like short column in shape or long shuttle-shape while paralleled. After four weeks, most cells became short column-shape and touched with adjacent cells tightly to form myotube-like structure,with apparent directionality of cell arraying. BMSCs induced by bFGF were identified by the positive staining for desmin, α-sarcomeric actin and C-TnT. Of BMSCs, there were more desmin positive and α-actin-positive cells made up higher of all BMSCs than C-TnT-positive cells. Desmin and α-actin-positive cells were about 33.82% and 58.64%, and C-TnT-positive cells were about 28.94%. Desmin(α-actin ) appeared red, and C-TnT was green under a laser scaning confocal microscopy. Their co-expression appeared yellow. Transmission electron microscope showed that the induced cells were rod in shape and the ovoid nuclei were positioned in the center of the cell. lots of mitochondria, rough endoplasmic reticulum, ribosome and paralleled myofilaments were founded in plasm.RT-PCR assessment showed that the differentiated cells began to express GATA-4 from day 7 to day 28 of differentiation and began to express α-myosin heavy china(α-MHC) fro Related Articles | Metrics

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Analysis of stanniocalcin-1 expression and plasma membrane Ca SUP>2+/SUP> -ATPase activity in enterocytes of toad EM>Bufo bufo gargarizans/EM> 2011, 42 (6):  782-786.  doi: 10.3969/j.issn.0529-1356.2011.06.013 Abstract ( )   Objective To investigate the relation between plasma membrane Ca SUP>2+/SUP>ATPase (PMCA) activity and stanniocalcin-1 (STC1) gene expression in enterocytes in toad, EM>Bufo bufo gargarizans/EM> .Methods Based on tolerance test in this study, 72 adult toads were randomly divided into a high calcium group ［tap water supplied with 0.1mol/L CaClSUB>2/SUB>), a low calcium group (tap water supplied with 0.03mol/L ethylene diamine tetraacetic acid(EDTA)］ and a control group (tap water). The small intestinal mucosa was excised on the pre-exposure and post-exposure 12, 24, 48, 72, 96, 120, 144 hours, respectively. Both plasma Ca2 level and PMCA activity were detected with colorimetry, and STC1 gene expression was analyzed with semi-quantitative RT-PCR. Results The results showed that: (1) After 0.1mol/L CaClSUB>2/SUB> exposure, the plasma calcium level increased at the 12th and 24th hour (EM>P/EM><0.05), restored to the normal level at the 48 th hour, and went up at the 72 th hour (EM>P/EM><0.05). Additionally, PMCA activity was enhanced and STC1 gene expression was up-regulated during 12th-48th hour after 0.1mol/L CaClSUB>2/SUB> exposure. (2) After 0.03 mol/L EDTA exposure, the plasma calcium level decreased, but both PMCA activity and STC1 gene expression did not change.Conclusion High plasma calcium level stimulates PMCA activity and STC1 expression in enterocytes, whereas up-regulated STC1 may inhibits PMCA activity i Related Articles | Metrics

肿瘤生物学

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Ad-hDCT (tumor vaccine) inhibiting the proliferation of intracranial B16 melanoma cells in C57BL/6 mice 2011, 42 (6):  787-791.  doi: 10.3969/j.issn.0529-1356.2011.06.014 Abstract ( )   Objective To observe inhibitive effect on the intracranial B16 melanoma cells in C57BL/6 mice by vaccination with Ad-hDCT. Methods Forty C57BL/6 mice were divided to 2 groups: Ad-BHG control group and Ad-hDCT treatment group. B16-F10 cells were planted into the brain by mouse stereotaxie apparatus. General observation, flow cytometry, paraffin sections with HE staining，frozen sections with CD8 and F4/80 immunohistochemistry detection were adopted in this study.Results Seven days after vaccination with Ad-hDCT, the level of hDCT specific CD8+INF-γ+ T cells increased significantly in the peripheral blood of the mice (P 0.01 vs Ad-BHG group). Compared to 100% endpoint of Ad-BHG blank vector control, the survival rate was 60% in Ad-hDCT immunized group after B16 cells plantation in parenchyma 16 days. The mice behaviour was normal. Histological analysis indicated that no tumor cells proliferated in the vaccinated mice. CD8+ T cells were detected in the tumor mass to recognize and kill tumor cells in vaccinated animals. Furthermore, much more microglia were found around the tumor area in the brain of the vaccinated mice.Conclusion Ad-hDCT can elicit a strong cytotoxic T lymphocyte (CTL) response in C57BL/6 mice, leading to protection against Related Articles | Metrics

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Effect of melatonin on proliferation inhibition and apoptosis induction in the murine foregastric carcinoma cell EM>in vivo/EM> and EM>in vitro/EM> 2011, 42 (6):  792-797.  doi: 10.3969/j.issn.0529-1356.2011.06.015 Abstract ( )   Objective To investigate the effect of melatonin(MLT) on proliferation inhibition and apoptosis induction in the murine foregastric carcinoma (MFC) cell EM> in vivo/EM> and EM>in vitro/EM> .Methods We performed an EM>in vivo/EM> study by inoculating MFC cell line in mice. The mice models were successfully established as follows: Group A. Normal control mice; Group B, Tumor-bearing control mice with daily intraperitoneal injection of 100 mg/kg saline water; Group C, Tumor-bearing mice with low dosage of 25 mg/kg MLT via intraperitoneal injection; Group D, Tumor-bearing mice with medium dosage of 50 mg/kg MLT; Group E, Tumor-bearing mice with high dosage of 100 mg/kg MLT. One week after MLT injection, tumor samples were collected and weighted. EM>In vitro/EM> experiment, we performed a cell model by MFC cells treatment with different concentrations of melatonin, and then the effect of MLT on MFC cell proliferation and cycle change was detected by using double immunofluorescence staining, flow cytometry, CCK-8 and other methods.Results Compared with the tumor-bearing control mice, the tumor weights and volumes of the melatonin treated tumor-bearing mice group were significantly reduced. Compared with the blank control, melatonin had inhibitory effect on gastric cancer cell proliferation EM>in vitro/EM> , increased cell apoptosis and arrested of the cell cycle at the G2/M phase, which exhibited a dose dependent.Conclusion Melatonin has a potent anti-gastric cancer proliferation effect EM>in vivo/EM> and EM>in vitro/EM> , and the mechanism is associated with concentration and time-dependent effect of melatonin-induced cell apoptosis. This apoptosis was also related to the arrest of the cell cycle at the G2/M phase EM>in vitro/EM> . Related Articles | Metrics

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Expression of annexinA5 in uterine cervical carcionma of different clinical stages 2011, 42 (6):  798-801.  doi: 10.3969/j.issn.0529-1356.2011.06.016 Abstract ( )   Objective To study expression of annexinA5 (ANXA5) in uterine cervical squamous carcinoma of different clinical stages in order to explore the possibility of using ANXA5 as a clinical stage marker. Methods According to the international FIGO methods, the carcinoma tissues were classified as stages Ⅰ, Ⅱ and Ⅲ. RT-PCR method was used to determine the ANXA5 mRNA expression; Western boltting method and immunohistochemistry were employed to determine the protein expression. Comparisons between any two groups were analyzed by t-test.Results According to RT-PCR, intensities for ANXA5 mRNA expression were lower at stage Ⅰ than at stages Ⅱ and Ⅲ（EM>P /EM><0.05）. According to Western blotting, a total amount of ANXA5 protein was Ⅰ lower at stage Ⅰ than at stages Ⅱ and Ⅲ（EM>P /EM><0.05）, whi Related Articles | Metrics

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Expression of LGR5 in hepatoma cells, HCC tissue and its significance 2011, 42 (6):  802-806.  doi: 10.3969/j.issn.0529-1356.2011.06.017 Abstract ( )   Objective To investigate the expression of LGR5 in human hepatocelluar cancer tissues and four human liver cancer cell lines and to evaluate its value in the development of hepatocellular cancer.Methods The mRNA and protein expression of LGR5 in human liver cancer cell lines and normal liver cell were detected by real-time PCR and Western blotting. Expression of LGR5 and β-catenin in hepatocellular cancer tissue was determined by immunohistochemistry. LGR5 localization and expression in HepG2 cells were tested by immunocytochemical staining. Results Western blotting and real-time PCR results demonstrated that the expression of LGR5 decreased significantly in HepG2, Hep3B, Huh7 and PLC cells compared to LO2 cells(EM>P/EM> <0.05). Immunohistochemistry results also showed that LGR5 was downregulated in cancer tissue compared to adjacent tumor tissue, whereas β-catenin expression was upregulated in cancer tissue. Conclusion Compared to adjacent tumor tissue and normal liver cells, LGR5 expression was significantly downregulated in 36 cases of hepatocelluar cancer tissues and four kinds of h Related Articles | Metrics

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Expression of leptin receptor in breast cancer and its correlation with breast cancer related genes 2011, 42 (6):  807-809.  doi: 10.3969/j.issn.0529-1356.2011.06.018 Abstract ( )   Objective To study the expression levels of leptin receptor（LEPR） in breast cancer cell lines, normal breast samples and malignant breast cancer samples, and to analyze the expression correlation between LEPR and estrogen receptor（ER）, E-cadherin（E-Cad）, c-Myc and cyclin D1 in order to provide mechanism clues for obesiety associated breast cancer. Methods Using RT-PCR and Western blotting, we examined the expression of LEPR in different breast cancer cell lines. The expression level of LEPR was analyzed by RT-PCR in 5 paired breast samples. In the meanwhile, The expression of LEPR, ER, E-Cad, c-Myc and cyclin D1 was analyzed using realtime PCR in 45 breast cancer samples and 15 normal breast samples. The correlation was analyzed between the expression of LEPR and the expression of ER, E-Cad, c-Myc and cyclin D1 using SPSS statistic software. Results LEPR mRNA and protein were expressed in all breast cancer cell lines tested in this study. The expression of LEPR in the breast cancer samples was higher than in the paired adjacent normal breast tissues. With the statistic analysis, the expression of LEPR was correlated with expression of ER in both normal and malignant breast samples. LEPR expression was also close correlated with E-Cad and c-Myc expression in the breast cancer samples, but not with cyclin D1 expression. Conclusion LEPR expresses in breast cancer cell lines. Compared to the normal breast tissues, the expression of LEPR in breast c Related Articles | Metrics

解剖学

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Comparison between anatomic and radiographic measurement of C2 after trans-lamina screwing 2011, 42 (6):  810-814.  doi: 10.3969/j.issn.0529-1356.2011.06.019 Abstract ( )   Objective To provide the anatomic and radiographic data, verify the clinical application of trans-lamina screws and evaluate the efficacy of our radiographic methods. Methods The anatomic and radiographic measurements of C2 vertebra were conducted on cadavers and living subjects. A total of 96 human adult cadavers and 112 volunteers without upper cervical abnormality were studied in this project. The minimal height (H1), thickness (T), length (L1) of C2 lamina, height of the root of lamina (H2), distance from the entry point to the lateral rim of lamina (L2) and to the lateral rim of lateral mass (L3) were bilaterally measured using calipers. The spino-laminar angles (angle A) were also included. All measurements were taken at the thinnest part of the lamina in the axial and coronal planes. Results The data of males were significantly higher than that of females (EM>P /EM><0.05) except angle A. There was no significant difference in the values of bilateral lamina between group A and group B (EM>P/EM> >0.05).The thickness of the vertebral lamina was less than 6mm in 45% of the specimens. The length of the lamina in all specimens was less than 2.5cm, while only 5% of the specimens had a >3cm distance from the entry point to the rim of lamina. The length from the entry point to the lateral rim of lateral mass ranged from 2.5cm to 4.6cm，with the length longer than 4cm was seen in 4% specimens. BR>Conclusion The preoperative radiographic evaluation is very important to determine the suitable size of screws. The radiographic measurement method we used is simple, accurate and reliable for preoperative measu Related Articles | Metrics

组织学胚胎学发育生物学

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Effects of diammonium glycyrrhizinate lipid ligand on NF-κB expression in the rat with nonalcoholic fatty liver disease 2011, 42 (6):  815-819.  doi: 10.3969/j.issn.0529-1356.2011.06.020 Abstract ( )   Objective To study therapeutic effects and underlying mechanisms of diammonium glycyrrhizinate lipid ligand (DGLL) on nonalcoholic fatty liver disease (NAFLD). Methods Sixty SD rats were randomly divided into six groups(10 rats per group): the model group, 3 DGLL-treated (30, 60, 120mg/kg) groups, the positive control ［treated with biphenyldicarboxylate (200mg/kg)］ group and normal control group. The stomach of the rat of the model group was irrigated with high lipid emulsion. The rats of DGLL groups were irrigated with relative dose of DGLL. After nine weeks, the changes of total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL), high density lipoprotein (HDL) in serum and the contents of malondialdehyde (MDA), superoxide dismutase (SOD) in liver homogenate were measured; the NF-κB p65 level in liver tissues was observed by immunohistochemistry, respectively. The expressions of NF-κB p65 and I-κBα in liver were determined by Western blotting. BR>Results Compared with the model group, the levels of TC，TG，LDL，MDA were decreased, meanwhile the activity of HDL and SOD were enhanced, the level of NF-κB in liver tissues was ameliorated remarkably by administration of DGLL. Meanwhile, th Related Articles | Metrics

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Effect of transplanted bone marrow mesenchymal stem cells on the repair of colon tissues of rats with ulcerative colitis 2011, 42 (6):  820-824.  doi: 10.3969/j.issn.0529-1356.2011.06.021 Abstract ( )   Objective To observe the effect of transplanted bone marrow mesenchymal stem cells(MSCs) on repairing of the colon tissues of rats with ulcerative colitis. Methods The MSCs from rats were isolated, cultured and labelled with fluor. Forty-five SD Rats were randomly grouped as A, B and C groups. Immune combined TNBS/ethanol was induced in groups A and B. Group C was the control group. The rat in group A received a caudal vein injection of 1ml of a fluids containing the labelled MSCs, and the rat in group B received 1ml of PBS after the induction of colitis. 5 rats in each group were sacrificed at day 3, 7 and 14 after injection. The colonic tissue in 3 groups were collected and prepared as paraffin sections for examination of pathological changes. Some colonic tissues in group A were prepared as frozen sections to observe the distribution of transplanted cells in the colon under a fluorescent microscope, and expression of cytokeratin(CK) of transplanted cells by immunofluorescent staining under a laser confocal microscope. Results The MSCs proliferated rapidly and had a typical fibroblastlike morphology. The repairing of injured colon in group A was much better than that in group B on day 7 and 14. The transplanted cells were seen in colon in group A, and the number of the cells increased as time going and was maximized on day 14(P <0.05). There were a few transplanted cells expressed CK in group A on day 7 and 14. Conclusion The transplanted MSCs distribute in the injured colon of rats with ulcerative colitis, and some MSCs diferentiate into colonic epithelial cells and participate in repair of the i Related Articles | Metrics

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Effect of diabetes on the differentiation and osteogenesis mineralization offemoral head growth plate in rats 2011, 42 (6):  825-831.  doi: 10.3969/j.issn.0529-1356.2011.06.022 Abstract ( )   Objective To study the effect of diabetes on development of long bones.Methods The diabetic model(STZ) of the rat was established.Rats were randomly divided into 3 normal groups：5-week(CON1), 10-week(CON2) and 15-week(CON3) and 3 diabetic groups：5-week(DM1), 10-week(DM2) and 15-week(DM3),10 rats each group. The change of collagen fiber depositing was observed by Van Gieson staining method. The microvascular density was measured by the ink infusion method.The techniques of in situ hybridization and RT-PCR were used in this study in order to analyze the intensity of vascular endothelial growth factor (VEGF)mRNA expression and content of decorin,respectively.Results The thickness and cellular density of the cell pillar in the growth plate were obviously lower in the 15-week diabetic group than those in the control group(EM>P/EM> <0.05).The content of collagen in the calcified zone of the growth plate was higher than that in the control group(EM>P/EM> <0.05),and the content of decorin decreased in the 15-week diabetic group(EM>P/EM> <0.01),the intensity of VEGFmRNA expression increased(EM>P/EM> <0.01),the microvascular density increased.Conclusion The cellular proliferation,differentiation and its osteogenesis mineralization in the growth plate of the femoral head in the 15-week diabetic rats a Related Articles | Metrics

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Expression of microtubule-associated protein 2 and nestin in the developing spinal cord of the human embryo 2011, 42 (6):  832-835.  doi: 10.3969/j.issn.0529-1356.2011.06.023 Abstract ( )   Objective To investigate distribution of the microtubule-associated protein 2 (MAP-2) and nestin in the human developing embryonic spinal cord. Methods Sixteen human embryos, ranged from 2 to 4 months old were used in the present study. Immunohistochemical staining was uesd to detect the expression and distribution of MAP-2 and nestin in the anterior horn, central canal and posterior horn of the spinal cord. Results At the stages of the second to fourth months, nestin-positive nerve fibers were seen within the human embryonic spinal cord, with increase of the gestational age, the expression of nestin-positive neural precursor cells and glial cells first increased and then decreased in the anterior horn, but that in the posterior horn gradually increased. MAP-2 in the anterior and posterior horns of spinal cord nerve cells were positive, with increasing gestational age. Conclusion MAP-2 and nestin involve in the regulation of spinal cord neurons and glial cells during the human embryonic growth and development. Related Articles | Metrics

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Proliferating cell nuclear antigen and c-Fos protein regulating development of the human embryonic spinal cord 2011, 42 (6):  836-839.  doi: 10.3969/j.issn.0529-1356.2011.06.024 Abstract ( )   Objective To explore effects of proliferating cell nuclear antigen (PCNA)and c-Fos protein on the early and middle embryonic development of the human spinal cord. Methods Using immunohistochemical method, the expression of PCNA and c-Fos were investigated in the central canal, anterior horn and posterior horn of spinal cord of 16 human embryos aged at the second to fourth month of gestation. Results At the second month of gestation, there were the positive PCNA immunohistochemical reaction in the central canal and alar plate of the spinal cord, but not in the basal plate. The c-Fos immunohistochemical positive reaction were localized in the epithelial cells of the central canal, alar plate and basal plate of the spinal cord. Following growth, the positive immunohistochemical staining for PCNA was also found in the epithelial cells of the central canal, anterior horn, and posterior horn of the spinal cord. The average intensity and cell number of the c-Fos immunohistochemical positive profiles decreased first, and then increased in spinal cord anterior horn (EM>P /EM><0.01), but no changes was found in the posterior horn of the spinal cord. Conclusion PCNA and c-Fos proteins may regulate the growth and development of the neuroepithelial cells of the neural tube of the human embryo. Related Articles | Metrics

人类学

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Variation of morphological traits in head and face of Han in Shanxi province 2011, 42 (6):  840-845.  doi: 10.3969/j.issn.0529-1356.2011.06.025 Abstract ( )   Objective To study changes of head-face morphological characters with the increasing age. Methods Thirty-eight head-face characteristics of 401 male (150 urban males and 251 rural males) and 402 female adults (153 urban females and 249 rural females) of Han were investigated in Qi county of Shanxi province in August 2009. Twelve physical indices were calculated. Preliminary analysis was carried out to determine head-face characteristics changes with the increasing age. Results The head breadth，minimum frontal breadth, face breadth, external biocular breadth，lip height，thickness of lips and auricular height had negative correlations with age. Head length, nasal breadth, mouth breadth, morphological facial height, nasal height, nasal length, nasal depth, upper lip skin height, physiognomic ear length, physiognomic ear breadth and facia skinfold had positive correlations with age. With increase of the age, Shanxi han nationality’s mongoloid fold rate and external angle rate dropped, the internal angle rate increased, the opening height of eyeslits became narrower, the brown eye rate increased, the black eye rate dropped. The rate of upper lip skin height increased ,but the middle rate and narrow rate decreased. The thin rate of thickness of lips increased obviously, but thick rate fell. The middle rate of breadth of alae nasi increased. Length-breadth index of head, length-height index of head lip index had negative correlations with age. Morphological facial index, and vertical cephalo-facial index had positive correlations with age.Conclusion Changes of the headface morphological charac Related Articles | Metrics

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Somatotype of urban adults of Han in Liaoning province by using Heath-Carter method 2011, 42 (6):  846-850.  doi: 10.3969/j.issn.0529-1356.2011.06.026 Abstract ( )   Objective To understand Liaoning urban adults of Han character size. Methods The body shape of 519 (men and women 257, 262) Liaoning urban adults were analyzed by using Heath-Carter method.Results The average size of Liaoning urban Han male adults was 4.7-4.2-1.6, in the case of endodermal-mesoderm and balanced body, and that of women was 6.4-3.5-1.6, mesomorphic endomorph. The gender differences of Liaoning urban Han adult body were as follows: the endomorphy of the females was much more than that of the males, the mesomorphy of the males was higher, therefore the males’ body was relatively strong, subcutaneous fat of femals was more developed. The body comparison between Liaoning Han urban adult and other groups showed that: Liaoning Han males figure was close to that of the Shandong Han,the United States Northeast and Mongolia, while the female figure was close to that of India, Mongolia and Shandong females. Conclusion Liaoning urban Chinese adults have a thick subcutaneous fat, bone, a more developed muscle system and the linearity of medium body. Male bones and muscles are in good shape.Female subcutaneous fat is well developed, iI>&#/I>8226;eI>&# Related Articles | Metrics

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Correlation between stature and arm span in young and middle aged adults of rural Han nationality in Liaoning 2011, 42 (6):  851-853.  doi: 10.3969/j.issn.0529-1356.2011.06.027 Abstract ( )   Objective To investigate the stature and arm span of young and middle aged adults of rural Han nationality in Liaoning. Methods A methodology and a standard of anthropology were used to measure arm span and stature of 769 young and middle aged adults (males 386, females 383) of rural Han nationality in Liaoning province. The original data were analyzed by medical statistics and regression analysis. Results Based on the data of the stature and arm span obtained in this study, the mean, difference, typing of differences, exponent and regression equations were determined for young and middle aged adults of rural Han nationality in Liaoning. Conclusion The measurement and typing among the stature and arm span have uniformity with Chinese stature and the difference of height and sex. The stature has high correlation with the arm span. It is reliable to use the regression equations to estimate the stature or the arm span. Related Articles | Metrics

生物工程学

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Custom design and primary construction of the mandibular condyle scaffold 2011, 42 (6):  854-858.  doi: 10.3969/j.issn.0529-1356.2011.06.028 Abstract ( )   Objective To explore the method of designing and manufacturing the mandibular condyle scaffold individually by rapid prototyping technologies and reverse engineering.Methods Cranial CT image data were processed in Mimics software for reconstruction of one side of ramus of mandible, and were inputted into Solidworks software to edit by the format of stl. A negative mold of the mandibular condyle scaffold was obtained. The mold was fabricated by resin materials utilizing rapid prototyping. Biomaterials were filled into the resin mold. When the materials were cured, we eliminated the resin mold and acquired a multi-pores three dimensional mandibular condyle scaffold model. The general morphology and microstructure of the scaffold model were observed by scanning electron microscopy (SEM). Results The mandibular condyle scaffold made of biomaterials was accordance with the one our computer designed. SEM observation revealed that the model was made of collagen in the cartilage-like layer and calcium phosphate cement/poly (lactic- co-glycolic acid) in the bone-like layer. Conclusion It is feasible to fabricate mandibular condyle scaffold individually by using reverse engineering and rapid prototyping technology. Related Articles | Metrics

技术方法

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A method of spatial coordinates conversion in the digital human brain 2011, 42 (6):  859-861.  doi: 10.3969/j.issn.0529-1356.2011.06.029 Abstract ( )   Objective To describe a spatial coordinates conversion method for serial anatomical cross-sectional images of the human brain using the software, Excel 2003.Methods Initialization reference frame was established in serial anatomical cross-sectional images of the human brain. Then, a standard spatial coordinates was set up by one time horizontal moving and 3 times rotations of the reference frame. Results A datasheet about coordinates conversion from initialization reference frame to standard spatial coordinates was established. Conclusion Standard spatial coordinates was established in the digital human brain after coordinates conversion. It will be helpful to build Chinese digital brain model. Related Articles | Metrics