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2012, Volume 43 Issue 3
06 June 2012 Previous Issue    Next Issue
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论文
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
技术方法


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论文

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A brief introduction to combined stem cell/gene therapy 2012, 43 (3):  289-292.  doi: 10.3969/j.issn.0529-1356.2012.03.001 Abstract ( )   The merge of cell therapy with gene therapy forms a more effective combined stem cell therapy/gene therapy. An introduction to combined stem cell therapy/gene therapy is made in this minireview briefly, including the often used cell types, vectors and their advantages and disadvantages. Examples of some diseases in human being or in animals which have been treated with combined stem cell therapy/gene therapy successfully are given also. It is pointed out that carcinogenesis might occur during the treatment and how to overcome such problems. Related Articles | Metrics

神经生物学

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Morphological changes of the nigrostriatal pathway in a mouse model of Parkinson’s disease 2012, 43 (3):  293-298.  doi: 10.3969/j.issn.0529-1356.2012.03.002 Abstract ( )   Objective To observe morphological changes in the nigrostriatal pathway by progress of parkinsonism in a mouse model, in order to investigate the pathophysiology of Parkinson’s disease from different contexts and to provide some new clues for Parkinson’s prevention, delay, and treatment.Methods Thirty-six C57 mice were randomly divided into the saline control and the1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) injection groups, with eighteen mice per group. The number of dopaminergic neurons in the substantia nigra, their nerve fiber densities and distributions on the striatal neurons with the direct and indirect pathway cells (dopamine D1 receptors/dopamine D2 receptors positive, D1R/D2R positive) in the striatum were observed by immunohistochemistry and confocal microscopy at 7, 14, 21, 25, 28 and 35 days after the injection.Results The number of dopaminergic neurons reduced to 40%-50% in the substantia nigra after injection of MPTP. Compared with the control, the decrease of dopaminergic fibers in the striatum showed a tendency that had an early reduction followed by an increase, with a minimum at day 21 (loss of 20% fibers), and reaching 45% fibers at the 35th day after injection. The distribution ratio of the dopaminergic terminals on striatal neurons to D1R/D2R positive cells was relatively low at day 21, and reached a peak on the day 35.Conclusion The dopaminergic fibers in the striatum show a significant regeneration after a MPTP lesion of the substantia nigra, and these regenerated fibers distribute mostly on those neurons in the direct pathway. The balance of dopamine’s activity regulation on the direct and indirect pathways of basal ganglia is broken. This may be an importa Related Articles | Metrics

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Autophagy in hippocampal neurons of sphingomyelin synthase 2 knockout mice 2012, 43 (3):  299-305.  doi: 10.3969/j.issn.0529-1356.2012.03.003 Abstract ( )   Objective To investigate the influence of ceramide on the autophagy of the hippocampal neurons of sphingomyelin synthase 2 knockout (SMS2SUP>-/-/SUP> ) mice. MethodsSMS2SUP>+/-/SUP>mice were bred by varied methods of hybridism, backcross and cross. The genomic DNA of these mice was achieved by the phenol chloroform extraction method. Then the target DNA was amplified by PCR method andidentified by agarose gel electrophoresis to establish the SMS2SUP>-/-/SUP>mice model. The autophagy of hippocampal neurons in P7, P14 and P30 in CA1 area of the SMS2SUP>-/-/SUP>mice was investigated by transmission electron microscopy, immunofluorescent labeling method and Western blotting technique with 8 mice per time group.Results 1.The autophagy phenomenon of hippocampal neurons of SMS2SUP>-/-/SUP>mice: With the electron microscopy, numerous lysosome and auto-phagosomes were found in the hippocampal neurons in SMS2SUP>-/-/SUP>mice model. Under the light microscope, the number of autophagy cells of P7, P14 and P30 in the CA1 area of the model were obviously higher than that of control group (EM>P/EM> <0.01); 2.Accommodation of Beclin-1 on autophagy phenomenon: The difference of the expression of Beclin-1 between model and control group was consistent with microtubule-associated protein 1 light chain 3 (MAPLC3). The number of positive cells of Beclin-1 in model group of P7, P14, P30 were obviously higher than that in control group (EM>P/EM> <0.01). The co-expression of Beclin-1 and MAPLC3 was found in the same hippocampal neurons; 3.The result of Western blotting analysis was consistent with that from immunocytochemistrys.Conclusion The number of ceramide in neurons of SMS2SUP>-/-/SUP>mice increases which not only promotes the apoptosis, but also promotes the autophagy of the neurons. Related Articles | Metrics

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Role of protease activated receptor-1 and 4 in the activation of microglia following subarachnoid hemorrhage 2012, 43 (3):  306-311.  doi: 10.3969/j.issn.0529-1356.2012.03.004 Abstract ( )   Objective To investigate the role of protease activated receptor-1 and 4 (PAR-1, PAR-4) in the activation of microglia following subarachnoid hemorrhage (SAH). Methods One hundred male SD rats were randomly divided into: sham, SAH+control siRNA, SAH+PAR-1 siRNA, SAH+PAR-4 siRNA and SAH+PAR-1/4 siRNA groups, 20 in each group. The drugs were administrated intracerebroventricularly at 1 hour after SAH. At the 6th and 24th hour after SAH, the neurological deficits, brain water content and Evans blue content were evaluated, at the same time, the expression of tumor necrosis factor α(TNF-α) and intercellular cell adhesion molecule-1 (ICAM-1) were also measured by using immunohistochemistry and Western blotting. Results PAR-1siRNA or PAR-4 siRNA could significantly ameliorate the neurological deficits and decrease vascular permeability (EM>P/EM>0.05). In addition, the expression of TNF-α and ICAM-1 was also decreased by PAR-1siRNA or PAR-4 siRNA treatment. The combination of PAR-1and PAR-4 siRNA played more powerful roles than either of them. Conclusion PAR-1 and 4 mediate the activation of microglia in the brain following SAH. Through suppressing the expression of inflammatory factors, PAR-1siRNA and/or PAR-4 s Related Articles | Metrics

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Fibroblast growth factor signaling regulates the delamination and migration of neural crest cells in the early chick embryos 2012, 43 (3):  312-316.  doi: 10.3969/j.issn.0529-1356.2012.03.005 Abstract ( )   Objective To investigate the effect of FGF on the migration of the neural crest cell in early chick embryos after blocking FGF signaling with Sprouty2 Methods The chick embryos were incubated EM>in vivo /EM>until HH9, Sprouty2 green fluorescent protein(GFP) plasmid was injected into the lumen of the neural tube using microinjection，and EM>in vivo/EM> electroporation was performed to transfect Sprouty2-GFP at a half-side of the neural tube while another half-side was used as the control side. We employed the immunofluorescence staining method to detect the HNK1-positive neural crest cell, specially labeled dorsal-ventrally, in order to determine whether Sprouty2 over-expression affects the delamination and migration of cranial and trunk neural crest cells. We studied if the FGF-related neural crest cell migration induced by Spourty2 transfection was controlled by regulating Ca+2-dependent N-Cadherin expression.Results The blocking FGF signaling via transfected Sprouty2-GFP resulted in more HNK1-positive neural crest cells in Sprouty2-GFP transfected side than in the control side in both cranial and trunk regions of the early chick embryos. The N-Cadherin expression did not significantly affected by Sprouty2-GFP transfection. BR>Conclusion The blocking FGF signaling using Sprouty2 promotes the neural crest cell delamination in the early chick embryos. However, N-Cadherin may not have the impact of FGF signaling on neural crest cell migration, on which Related Articles | Metrics

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Development of the rostral migratory stream and its cell apoptosis 2012, 43 (3):  317-317*321.  doi: 10.3969/j.issn.0529-1356.2012.03.006 Abstract ( )   Objective To study the development of the rostral migratory stream (RMS) of mice and the proliferation and apoptosis of neural stem cells in RMS at various postnatal ages.Methods With 5-bromodeoxyuridine (BrdU) assay and Caspase-8 immunofluorescent staining, the neuroproliferation and cell apoptosis in RMS were investigated.Results In early postnatal phase, there were many proliferative cells in the brain, especially in subventricular zone (SVZ) and RMS. With age increasing, the stem cells in the brain decreased. In the adult, the stem cells in the neocortex were hardly found except in SVZ and RMS areas. When the proliferation of stem cells was companied with apoptosis during RMS development, there was a positive correlation between cell proliferation and apoptosis.Conclusion There is an important signification for the neuroproliferation and cell apoptosis in RMS. Through cell apoptosis, the migration and differentiation of neural stem cells in RMS are regulated. Related Articles | Metrics

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Effects of Ghrelin on spatial memory, hippocampal neuron synapses and growth hormone secretagogue receptor expression in mice 2012, 43 (3):  322-327.  doi: 10.3969/j.issn.0529-1356.2012.03.007 Abstract ( )   Objective To investigate effects of Ghrelin on learning and memory processes, hippocampal neuron synapses and growth hormone secretagogue receptor (GHS-R) expression in mice. Methods Twenty male mice (8 weeks old) were randomly divided into the Ghrelin-treated and control groups (10 mice/group) in which Ghrelin or equal quantity saline was intraperitoneally injected, respectively. The ability of spatial learning and memory were tested with Morris water maze (MWM), expression of Ghrelin receptor GHS-R in hippocampus with immunohistochemical method and image analysis, and structural changes of synapsis in the hippocampal region by transmission electron microscope.Results In the MWM experiment, compared with the control group, the escape latency of the treated group on the day 2, 3 and 4, was significantly shortened, the times crossing hidden platform markedly increased (EM>P/EM> <0.05),expression of GHS-R immunoreactive products increased obviously in hippocampus (EM>P/EM> <0.05),and numbers of hippocampal neuron synapses in the CA3 region were also strikingly increased in the Ghrelin-injected group compared with the control group (EM>P/EM> <0.05).Conclusion Ghrelin may increase the synapse numbers of the hippocampal region, by binding the GHS-R in hippocampus, and subsequently enhance spatial learning and memory Related Articles | Metrics

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Therapeutic effects of lithium chloride on the elevated plus-maze behavior in FMR1 knockout mice 2012, 43 (3):  328-334.  doi: 10.3969/j.issn.0529-1356.2012.03.008 Abstract ( )   Objective To study the intervention and mechanism of glycogen synthase kinase 3 (GSK3) inhibitor, lithium chloride, on the elevated plus-maze behavior of fragile X mental retardation 1 (FMR1) knockout mice. Methods Ninety 30-days-old FMR1 knockout (KO) mice were used in this study and lithium chloride was intraperitoneally and continuously injected for 5 days. Elevated plus-maze test was processed in day 6. We monitored the trace of the mice and analyzed it with a software named Smart, to observe whether lithium chloride could ameliorate the phenotype of KO mice in elevated plus-maze. We also tested the expression of GSK3β and p-GSK3β in both cortex and hippocampus of KO and wild type (WT) mice with Western blotting. Results In the elevated plus-maze, we observed that, comparing with WT mice, KO mice had significantly higher moveability, excitability and exploratory. KO mice also had more time spent in open arms as well as the entry and total distance than WT mice. After lithium chloride administration, the time spent, entry and total distance of KO mice in the open arms had significantly reduction (EM>P/EM> <0.05). In Western blotting test, we found the expression of p-GSK3β of KO mice was less than WT mice. After treated with lithium chloride, KO mice had the increased p-GSK3β expression. WT mice had amelioration in time spent, entry and total distance in the open arms after lithium chloride applied. We observed the increased p-GSK3β expression in the lithium chloride treated WT mice. Compared the KO control with the WT control, there was no significant difference in total GSK3βexpression.After lithium chloride administration, there were no significance changes in total GSK3β expression (P >0.05).Conclusion The function of lithium which ameliorates elevated plus-maze behaviors Related Articles | Metrics

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Effect of bone morphogenetic protein-7 on expression of Caspase-3 in rats with cerebral ischemic reperfusion injury 2012, 43 (3):  335-339.  doi: 10.3969/j.issn.0529-1356.2012.03.009 Abstract ( )   Objective To explore the effect and the mechanism of bone morphogenetic protein-7 (BMP-7) on neuronal apoptosis in the rat brain after cerebral ischemic reperfusion injury. Methods Ten rats from 40 adult healthy male Sprague-Dawley rats received sham-operation, and the other 30 rats were subjected to middle cerebral artery occlusion (MCAO) for 2 hours, of which the 20 successfully modeled rats were equally randomized into the control group and the treatment group, 10 rats each group. The rats in the treatment group were intervened with 250μl of BMP-7 (0.1mg/kg) via caudal vein injection, while the rats in the control group and the sham-operative group were intervened with equal volume of normal saline. Animals were sacrificed at the 24th hour after reperfusion. Neurological deficits were evaluated by Bederson method, and the infarction volume of brain was investigated by 2,3,5-triphenyl tetrazolium chloride coloring. Pathological changes in the lesion site were investigated by HE coloring. The neuronal apoptosis was detected by TUNEL staining and the expression of Caspase-3 with the immunohistochemistry method.Results In the treatment group, the Bederson score (1.7±0.5, EM>t /EM>= 4.66, EM>P/EM> 0.01) and focus infarction ［(7.6±1.4）%, EM>t/EM> =6.98, EM>P /EM>0.01］ were lower than those in the control group ［2.7±0.5, (22.3±4.5)%］. Damage of the ischemia brain was significantly lessened in the BMP-7 treatment group. The number of TUNEL positive cells in the cortex (3.6±0.6), striatum (7.4 ±1.1) and hippocampus (5.0±0.7) of the treatment group were lower than those in the control group (13.4±1.1,17.8±1.5,15.4±1.1,EM> P/EM Related Articles | Metrics

细胞和分子生物学

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Role of growth associated protein-43 on regulating the cell cycle via calmodulin 2012, 43 (3):  340-346.  doi: 10.3969/j.issn.0529-1356.2012.03.010 Abstract ( )   Objective To explore the role of growth association protein-43 (GAP-43) in regulation of cell cycle via the interaction with calmodulin (CaM).Methods A series of NIH3T3 cell models, which expressed different types of GAP-43, i.e., 3T3-G (expression of wild-type GAP-43), 3T3-A (expression of non-phosphorylated GAP-43), and 3T3-D (expression of phosphorylated GAP-43), were established by using a retroviral expression system. A separate group of NIH3T3 cells, which was transfected only with an empty vector, were used as a control. Immunofluorescence and Western blotting were used for detecting expression of individual protein in transfected NIH3T3 cells and the co-immunoprecipitation was applied to detect the interaction of different types of GAP-43 with CaM. The cell counting, cumulative BrdU labelling, and the percentage of labeled mitotic figures (PLM labeling) were introduced to determine the cell cycle in different cell models.Results The cell models expressing different types of GAP-43 were successfully created and confirmed by immunocytochemistry and Western blotting. The co-immunoprecipitation showed a clear interaction between GAP-43 and CaM in 3T3-A cells, a little in 3T3-G cells, and no such interaction in either 3T3-D or 3T3-P cells. No significant change of cell cycle was observed by cumulative BrdU measurement,pulse BrdU test and M phase test in 3T3-P cells, while certain extensions were seen in 3T3-D and 3T3-G cells. In 3T3-A, the cell cycle was shown to be prolonged with an extension of G1phase.Conclusion The proliferation of NIH3T3 cells, caused by GAP-43 expression may be due to an interaction between GAP-43 and CaM, which may reduce binding capacity of CaM to the Ca SUP>2 +/SUP>, causing an extension of cell cycle, especially for Related Articles | Metrics

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Effects of rhEndostatin on the morphology and proliferation of human keloid-derived fibroblasts EM>in vitro/EM> 2012, 43 (3):  347-351.  doi: 10.3969/j.issn.0529-1356.2012.03.011 Abstract ( )   Objective To analyze the inhibitory effect of recombinant human endostatin(rhEndostatin) on human keloid-derived fibroblasts(KFs) in vitro.Methods The human’s KFs were cultured and identified in vitro. KFs were incubated with various concentrations of rhEndostatin solution(6.25×10-6, 1.25×10-5, 2.50×10-5, 5.00×10-5, 1.00×10-4mg/L).With the 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenny- BR>lterazolium bromide(MTT) colorimetric assay the effects of rhEndostatin on the proliferation of cultured KFs were studied,and morphological changes of the KFs were also observed at different time points(24, 48, 72 hours). Results KFs from the keloid tissue started to appear on days 7-8 after it were planted, the number of the cells gradually increased and the growth of the cells showed a radial patten centred at the tissue-centric. Second subculture KFs had a single-layer arrangement, were uniform, showed long fusiform, and had good refractive index.Mouse anti-Vimentin was positive.Buffy positive reaction product was observed in the cytoplasm. Cells growth inhibition occurred at 1.25×10-5mg/L or higher concentration in a dose-dependent manner and maximum inhibition occurred at 48 hours. After incubation with rhEndostatin(5.00×10-5mg/L and 1.00×10-4 mg/L) for 48 hours ,observation under the microscope showed the cells slowed growth, Related Articles | Metrics

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Comparative analysis of the role of JNK signaling pathway in cell proliferation and apoptosis of rat liver regeneration and cirrhosis 2012, 43 (3):  352-358.  doi: 10.3969/j.issn.0529-1356.2012.03.012 Abstract ( )   Objective JNK signaling pathway regulates some physiological activities, including inflammation, cell proliferation, differentiation, apoptosis and stress response. The study was to compare the role of JNK signaling pathway in rat liver regeneration (LR) and in rat liver cirrhosis (LC) at the gene transcription level.Methods The rat LR model was established by 2/3 partial hepatectomy(PH), and the LC model was induced by intraperitoneal injection of CClSUB>4/SUB> and colza oil. Rat Genome 230 2.0 array was used to detect gene expression profiles of JNK signaling pathway-related genes , and bioinformatics and systems biology methods were used to analyze the physical activities which were predicted by the expression profiles.Results JNK signaling pathway includes 302 genes and 42 branches. Of them, 240 genes were contained in Rat Genome 230 2.0 array, including 79 significantly expressed genes. In detail, 52 genes were the LR-specific, 5 genes the LC-specific, and 22 genes the common. The cell proliferation-promoting effect of paths 1 and 16 and the cell apoptosis-promoting effect of paths 22-33 became stronger at the priming phase of LR than in control; the cell proliferation-promoting effect of paths 1-17, 34 and 35 and cell apoptosis-promoting effect of paths 22-33 was enhanced at the progressing phase of LR, while the cell apoptosis-inhibiting effect of paths 37-41 and the cell apoptosis-inducing effect of paths 37, 39, 41 and 42 was weakened at the terminal phase. Paths 18-21 and 36 were found not to be involved in rat LR. JNK signaling pathway of LC worked as that of the control did. Conclusion Thirty-seven branches of the JNK signaling pathway regulat cell proliferation and apoptosis in rat LR, but may not be significantly involved in rat LC. Related Articles | Metrics

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Caspase signaling pathway regulates hepatocyte apoptosis in rat liver regeneration 2012, 43 (3):  359-365.  doi: 10.3969/j.issn.0529-1356.2012.03.013 Abstract ( )   Objective To explore the role of Caspase signaling pathway in regulating hepatocytes of the rat regenerating liver. Methods A total of 114 adult rats were randomly divided into 19 groups which consist of 9 partial hepatectomy groups,9 sham operation groups and 1 normal control group.Hepatocytes were separated with a conventional two-step perfusion method and the percoll density gradient centrifugation at 10 different time points following operation.Rat Genome 230 2.0 array was used to detect expression changes of Caspase signaling pathway-related genes in rat liver regeneration (LR) .Fluorescence quantitative PCR (FQ-PCR) was used to confirm reliability of microarray results.Bioinformatics and systems biology methods were used to analyze the role of Caspase signaling pathway in rat LR. Results Literatures showed that 38 paths and 123 genes were involved in Caspase signaling pathway.Of them,106 genes were included in Rat Genome 230 2.0 array,and it was found that 38 genes were related to hepatocytes of rat regenerating liver.A total of 21 paths from Caspase signaling pathway,including paths 1,2 and 11 at 30 hours,paths 27,29 and 31 at 70 hours,paths 15 and 16 at 2 hours and 30 hours,paths 3 and 4 at 30 hours and 72 hours after partial hepatectomy (PH),were found to inhibit hepatocyte apoptosis while 14 paths,such path 14 at 2 hours,path 34 at 6 hours,path 7 at 30 hours,path 13 at 2 hours and 30 hours,path 36 at 36 hours and 30 hours after PH,were involved in promoting hepatocyte apoptosis during rat LR. Conclusion Thirty-eight genes and 15 paths of Caspase signaling pathway may regulate apoptosis of hepatocytes in rat LR. Related Articles | Metrics

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Different appearance of senescence and apoptosis during primary culture of the mouse adipose-derived mesenchymal stem cellsEM> in vitro/EM> 2012, 43 (3):  366-371.  doi: 10.3969/j.issn.0529-1356.2012.03.014 Abstract ( )   Objective To investigate senescence and apoptosis of the mouse adipose-derived mesenchymal stem cells (ADMSCs) during primary culture EM>in vitro/EM> Methods ADMSCs of inguinal adipose tissue of one-month-old and 24-month-old mice, 30 of per group respectivly, were isolated and cultured primarily by differential adhesion methods. Expressions of CD73,CD90 and CD105 of P3 ADMSCs were detected by immunofluorescence technique and obsered under an fluoresence microscope. Morphology and growth status of ADMSCs were compared between two groups. β-galactosidase (SA-β-Gal) staining was used to identify the aging change of ADMSCs. The apoptosis of ADMSCs was detected by PI-Hocchst33342 double staining. Results The ADMSCs of two age groups cultured primarily in vitro were all adhered to grow and they showed the typical morphological characteristics of stem cells. The phenotypes of two age groups were ADMSCs, CD73, CD90 and CD105, positive. SA-β-Gal staining showed that proportion of the primary cultured aging ADMSCs in 24- month-old group was higher than those in the one-month-old group and higher than the primary cultured 2-7days ADMSCs in one-month-old group and in 24-month-old group were respectively. PI-Hoechst 33342 double staining showed that apoptotic proportions of primary cultured 2-7days ADMSCs in one-month-old group were (22.1±1.4)%, (36.7±1.62)%, (11.7±1.45)%, (38.4±1.57)%, (6.5±1.32)% and (11.3±1.63)%, respectively. Apoptotic proportions of primary cultured 4-7days ADMSCs in 24-month-old group were (29.4±1.72)%, (37.6±1.64)%，(19.4±1.29)% and (18.9±1.25)%, respectively. The number of apoptosis and dead cells had no significant difference. Conclusion The proliferation capability of primary cultured ageing ADMSCs is lower than that from the young age. Proportions of s Related Articles | Metrics

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Expression and purification of functional peptide Hepc25 of mouse Hepcidin 1 2012, 43 (3):  372-376.  doi: 10.3969/j.issn.0529-1356.2012.03.015 Abstract ( )   Objective To purify functional peptide Hepc25 of mouse and examine its expression by using the expression vector of prokaryote. Methods The gene of Hepc25 was fused with thioredoxin gene and the fusion protein was expressed greatly in E.ciol.Hepc25 gene amplification was done by RT-PCR and the gene was cloned into pET32a(+) plasmids. The pET-32a-Hepc25 plasmids were transformed into E.ciol BL21 competent cells.In order to obtain much of the fusion protein,the induction condition was optimized.The fusion protein was measured by SDS-PAGE and Western blotting.Isolation and purification of fusion protein were done by affinity chromatograph,iron exchange chromatography and exclusion chromatography.Purified Hepc25 from fusion protein hydrolyzed by enterokinase was analyzed by HPLC. Results BR>Fusion protein expression vectors were constructed successfully and the quantity of fusion protein was about 28% of total protein.Under the condition of 33℃,120 r/min,0.2 mmol/L isopropyl β-D-thiogalactopy ranoside(IPTG) for 8 hours,the quality of expression of fusion protein was the largest and the purity of fusion protein and Hepc25 were both over 95%. Conclusion Functional peptide Hepc25 of mouse is expressed by using prokaryotic expression vector and is purified.The optima exp Related Articles | Metrics

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Cathepsin B-RNAi-lentivirusd inhibitis capillary-like tube formationEM> in vitro/EM> 2012, 43 (3):  377-380.  doi: 10.3969/j.issn.0529-1356.2012.03.016 Abstract ( )   Objective To study inhibition of capillary-like formation by Cathepsin B-RNAi-Lentivirus (CTSB-RNAi-LV) EM>in vitro/EM>. Methods The control group and RNAi lentiviral group, and negative control group were tested in this study. Immunohistochemistry method and building capillary-like formation model in vitro were used to study the CTSB-RNAi-LV mouse brain microvascular endothelial cells (bEND.3), and explore affection of Cathepsin B and capillary-like formation. Results BR>Cell immunohistochemistry showed that Cathepsin B positive profiles appeared brow and were mainly located around the nucleus of bEND.3 cells. Of three groups, expression of Cathepsin B was the least in RNAi lentiviral group. Evaluation of bEND.3 the total protein and capillary-like formation showed that RNAi lentiviral group had less total protein content and formation of capillary-like zone than other groups and there was statistically significant (EM>P /EM><0.05).Conclusion CTSB –RNAi-LV can inhibit expression of Cathepsin B in the bEND.3 cell and formation of capillary-like zone in vitro. CTSB-RNAi-LV can inhibited targeted capillary-like formation EM> Related Articles | Metrics

肿瘤生物学

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Anticancer activity of the essential oil from Cinnamomum longepaniculatum leaves and its major components against human BEL-7402 2012, 43 (3):  381-386.  doi: 10.3969/j.issn.0529-1356.2012.03.017 Abstract ( )   Objective In order to study the anticancer activity of essential oil from Cinnamomum longepaniculatum leaves and its major components(1,8-cineole, sassafras oil, γ-terpinene, α-terpinene and terpinene-4-alcohol) against human BEL-7402 ceells. Methods By 3-4,5-diemethylthiarol-2,5-diphenytetrazolium bromide (MTT) fast staining method to study the growth inhibition rate of the essential oil from Cinnamomum longepaniculatum leaves and its major components on human BEL-7402 cells, and by soft agar cloning formation assay to study the effect of the essential oil from Cinnamomum longepaniculatum leaves and its major components against human BEL-7402 cells. Results The growth inhibition rates of the essential oil from Cinnamomum longepaniculatum leaves, 1,8-cineole, sassafras oil, γ-terpinene, α-terpinene and terpinene-4-alcohol against the BEL-7402 cells were 58.63%, 42.83%, 90.04%, 81.53%, 34.83% and 31.46%, respectively in vitro. Colony forming rate of the essential oil from Cinnamomum longepaniculatum leaves, 1,8-cineole, sassafras oil, γ-terpinene. α-terpinene and terpinen-4-alcohol on BEL-7402 cells were 55.13%, 61.99%, 94.56%, 89.54%, 62.02% and 59.91%, respectively. Conclusion The essential oil from Cinnamomum longepaniculatum leaves and its major components can inhibit the proliferation of live Related Articles | Metrics

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Influence of AGP on proliferation and expression of nuclear transcription factor p53 and c-myc in human breast cancer cell line MDA-MB-231 2012, 43 (3):  387-392.  doi: 10.3969/j.issn.0529-1356.2012.03.018 Abstract ( )   Objective To study the effects of acanthopanax giraldii harms var hispidus hoo polysaccharides (AGP) on proliferation and expression of nuclear transcription factor p53 and c-myc in human breast cancer cell line MDA-MB-231. Methods The proliferation rate of human breast cancer cell line MDA-MB-231, which was incubated by different concentration of AGP (0.15625, 0.3125, 0.625, 1.25, 2.5g/L) in DMEM medium for 1, 3, 5, 7 days in vitro respectively, was studied by WST-1 cell proliferation and cytotoxicity assay kit. The changes of human breast cancer cell line MDA-MB-231 morphology were observed by an inverted microscopy. The expression of apoptosis-related proteins such as p53 and c-myc was detected by fluorescence immunoassay and Western blotting. Results WST-1 assay showed that different concentration of AGP inhibited proliferation of human breast cancer cell line MDA-MB-231 treated for 3day, 5day, 7day when compared with the negative control group (P <0.05). The inhibition rate depended on AGP dose and reaction time. According to fluorescence immunoassay and Western blotting analysis, c-myc expression in cytoplasm reduced and p53 expression increased significantly after the treatment of AGP on human breast cancer cell line MDA-MB-231. Conclusion AGP has the effect of inhibiting proliferation on human breast cancer cell line MDA-MB-231.Its role may be related to reduce c-myc expression and increase p53 expression of human breast cancer cell line MDA-MB-231. Related Articles | Metrics

解剖学

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Angiographic investigation on venosomes and their anastomosis 2012, 43 (3):  393-397.  doi: 10.3969/j.issn.0529-1356.2012.03.019 Abstract ( )   Objective To investigate the method and mechanism of the venous angiography. Methods Forty SD rats were divided into two groups (A, B), 20 rats for each. Each group then was divided into 4 subgroups (a, b, c, and d). In group A, the rats were perfused with a contrast medium of various concentrations simultaneously through jugular vein and tail vein, whereas the rats in group B were perfused with a contrast medium of various concentrations through the carotid artery. All the rats in both groups underwent radiography simultaneously. The images obtained were processed with Photoshop CS4 to analyze the effects of venography. Skins of 4 New Zealand rabbits underwent radiography after perfusion with the contrast medium to investigate the mechanism of the venous drainage. Results The course and branches of a vein were clearly demonstrated in subgroup Ac, whose venography was achieved with 7% gelatin as the solvent. The accompaniment of the vein and the artery, and their relationship were clearly demonstrated in subgroup Bb, whose systematic arteriography and venography were achieved with 2% gelatin as the solvent. With sufficient pressure and perfusate amount, the main trunks and the communicating branches with a large-diameter in skins of the rabbits were clearly observed. Conclusion Not only the venous perfusion can be performed alone to achieve venography, but also the arterial and venous perfusion be performed at the same time to achieve excellent arteriography and venography simultaneously. To accomplish high-quality venography, the “bypass route” Related Articles | Metrics

组织学胚胎学发育生物学

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Effects of ephedrine on structure and polypeptides in the liver of filial mice BR> 2012, 43 (3):  398-404.  doi: 10.3969/j.issn.0529-1356. 2012.03.020 Abstract ( )   Objective To investigate changes of structures, expression of Bax protein activities of superoxide dismutase(SOD) and catalase(CAT), and contents of maleic dialdehyde(MDA) in the liver of filial mice and activity of glutamic-oxalacetic transaminease(GOT) and glutamic-pyruvic transaminase(GPT) in serum after injected ephedrine. Methods Totally 48 filial mice were used in this study. They were intraperitoneally injected with escalation doses (2.0g/L,3.0g/L and 4.0g/L ) ephedrine and the same amount of saline. Changes of the liver weight were examined. Activities of SOD, CAT, GOT, GPT as well as content of MDA were determined by colorimetry. Hepatic structures of 5, 10 and 15 days filial mice were observed by optical microscopy after injected ephedrine and the expression of Bax protein was measured by immunohistochemical staining. Results The liver weight of filial mice of the experimental groups was lower than the control groups at day 5 10 and 15 after injected ephedrine. The activity of SOD and CAT increased in the early ages and，then decreased with the extension of time and the increasing doses of the drug, SOD and CAT activities of the experimental groups were higher at day 5(EM>P/EM> <0.05） and lower at day 10(EM>P /EM><0.05） and day 15(EM>P/EM> <0.01） than that of the control group. While MDA content decreased firstly and then increased, MDA content of the experimental group lower than the control group at day 5 (EM>P/EM> <0.05), and higher than the control group at day 10 and 15(EM>P/EM> <0.01). The activity of GOT and GPT in filial mice serum of the experimental groups were higher than the control groups after injection of ephedrine(EM>P/EM> <0.05 orEM> P/EM> <0.01). Liver of filial mice appeared various degrees of damage after ephedrine injection, such as hepatic plate atrophy, hepatic sinusoid expand, the cell boundaries fuzzy and endotheliocyte fall off. The intensity of the expression of Bax protein of the experimental groups was higher than the control group after ephedrine injection(EM>P /EM><0.05 orEM> P/EM> <0.01). Conclusion The ephedrine affects the liver of developmental filial mice. This damage may be correlated with the lo Related Articles | Metrics

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Expression patterns of hyperpolarization-activated cyclic nucleotide-gated cation 4, connexin 43 and podoplanin in the embryonic mouse heart and development of cardiac conduction system 2012, 43 (3):  405-411.  doi: 10.3969/j.issn.0529-1356.2012.03.021 Abstract ( )   Objective To investigate the development of the cardiac conduction system of embryonic mouse heart. Methods Serial transverse sections of forty mouse embryonic hearts from embryonic day (ED) 9 to ED 16 were stained immunohistochemically or immunofluorescently with antibodies against hyperpolarization-activated cyclic nucleotide-gated cation 4 (HCN4), myosin heavy chain (MHC), connexin 43(CX43) and podoplanin. Results At ED9, strong positive expression of HCN4 was concentrated in the MHC negative sinus venosus, then gradually shifted to the sinoatrial node during the development of the embryonic heart. From ED11 onward, CX43 negative expression showed site-specific patterns. CX43 negative sinoatrial node was seen extending along the dorsal right atrial wall and left and right venous valves towards the dorsal wall of atrioventricular canal. At ED13, with fusion of base of the CX43 negative septum primum, left and right venous valves continued with the developing atrioventricular node derived from the dorsal wall of the atrioventricular canal. Developing atrioventricular bundle on the top of the interventricular septum was found connecting with atrioventricular node. From ED9 to ED10, strong positive expression of podoplanin was detected both in MHC positive myocardium and in myocardial precursors of splanchnic mesoderm of dorsal pericardial wall and mesenchyme surrounding sinus venosus. Between ED11 and ED13, podoplanin positive mesenchymal cells spread along outer surface of the heart and formed a podoplanin positive mesothelial epicardium. Conclusion At early stage of heart development, the leading pacemaker is located in sinus venous that exhibits function as a pacemaker earlier than contraction. CX43-negative myocardium observed at ED11 represents the establishment of the prototype of the developing cardiac conduction system. Podoplanin is involved in promoting myocardial precursor cells differentiating into cardiocytes. Related Articles | Metrics

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Effects of formaldehyde on the endogenous metabolism in the body 2012, 43 (3):  412-416.  doi: 10.3969/j.issn.0529-1356.2012.03.022 Abstract ( )   Objective To investigate effects of formaldehyde on endogenous metabolites of urine. Methods The urinary samples of the formaldehyde contact group and control group were analyzed by UPLC –Q-TOF/MS, combined with principal component analysis(PCA). The overall disease pattern of the metabolic network was established based on metabolic profiles in different groups. Results The different trajectory of urine was visually noted from metabolic profiles between 2 groups. Metabolomics appeared to be suitable for toxicity studies of formaldehyde, and a biomarker structure was identified. Conclusion The science connotation of the toxicity of formaldehyde may be elucidated by Metabonomics, according to metabolites variance in certain disease Related Articles | Metrics

人类学

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Correlation between basic fibroblast growth factor gene single nucleotide polymorphisms and osteoporosis in male of Zhuang nationality in Guangxi Baise 2012, 43 (3):  417-421.  doi: 10.3969/j.issn.0529-1356.2012.03.023 Abstract ( )   Objective To investigate the correlation of the basic fibroblast growth factor (bFGF) gene 5 single nucleotide polymorphisms(SNP) and bone mineral density(BMD) in Guangxi Zhuang nationality men. Methods A case-control study was carried out on 147 osteopenia patients and 154 matched controls. Inclusion criteria were at the age of 41 to 78, Zhuang nationality,living in Baise more than 10 years, and no history of taking drugs affecting bone metabolism. Exclusion criteria included secondary osteoporosis, any disease affecting bone mineral density or consanguinity. Genotypes for adiponectin gene 5 locis (rs308379、rs12644427、rs3789138、rs308442 and rs3747676) polymorphism were determined by Multiplex SNaPshot. Broadband ultrasound attenuation (BUA) for the right leg calcaneal was measured by French production of Osteospace dry ultrasound bone densitometer. Results rs308379、rs12644427、rs3789138、rs308442 and rs3747676 polymorphisms were met with HardyWeinberg equilibrium ( EM>P/EM> >0.05). Five locis genotype frequencies in the normal group and the osteopenia group were met with Hardy-Weinberg equilibrium (EM> P/EM> >0.05). There was no significant difference in the genotype distributions of five locis polymorphism between LBM group and control group ( P >0.05). Multivariate Logistic regression analyses revealed only rs308442 polymorphism remained significantly associated with low bone mass(LBM) ( EM>P/EM> <0.05). The subjects with the combined TA genotype had higher risk of LBM compared with those with the TT genotype( EM>OR/EM> =1.831,95% EM>CI/EM> :1.075-3.116,EM> P/EM> =0.026).Polymorphism of rs308442 was independently correlated with LBM at the calcaneal in Baise Zhuang femail population. Conclusion bFGF gene intron 1 rs308442 polymorphism and BMD in Baise Prefecture Zhuang men have some relevance. The presence of the T allele A may dominantly increase the risk of osteopenia. TT genotype has a protective effect of BMD.The data also suggest th Related Articles | Metrics

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Effects of apoE polymorphism on serum apoA1 and apoB levels in Maonan ethnic minority 2012, 43 (3):  422-424.  doi: 10.3969/j.issn.0529-1356.2012.03.024 Abstract ( )   Objective To investigate effects of apolipoprotein (apo) E polymorphism on serum apoA1 and apoB levels in Maonan ethnic group. Methods Serum apoA1 and apoB concentrations were measured in 221 healthy subjects from a population of Maonan ethnic group in Qiannan autonomous area of Guizhou province using immunoturbidimetric assay. ApoE2,E3,E4 genotypes were determined by polymerase chain reactionrestriction fragment length polymorphisms (PCR-RFLPs). Results Increased apoB levels and decreased apoA1-to-apoB ratio were found in carriers with apoE4 isoform (E3/4+E4/4) ( n =41) and apoE3/3 homozygotes ( n =143) as compared to carriers with apoE2 isoform (E2/2+E2/3+ E2/4) ( n =37, P <0.05). ApoB levels were higher in carriers with apoE4 isoform than those with apoE3/3 homozygotes ( P <0.05). No significant differences were found for apoA1 levels among the groups mentioned above ( P >0.05). Conclusion This study suggests that apoE polymorphism is associated with serum apoB levels and apoA1/B ratio, but not with apoA1 levels, in Maonan e Related Articles | Metrics

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Physical anthropology of the Miao nationality of Xishui in Guizhou 2012, 43 (3):  425-429.  doi: 10.3969/j.issn.0529-1356.2012.03.025 Abstract ( )   Objective To compare the traits on the head and face constitution between males and females of Miao Nationality in Xishui, and to identify the kinship and difference between the above group and the other 5 minority groups living in southern China and other groups living in Guizhou nationalities. Methods An assessment of the somatoscopy and anthropometry traits on the head and face (fourteen observations and nineteen measurements) of 357 Miao nationality adults aged from 20 to 59 years old (173 males and 184 females) living in Liangcun township of Xishui city in Guizhou was carried out and the statistical software was used to process data and cluster analysis. Results Of Xishui Miao facial features, there were significant sex differences on the nose breadth, nose height, nasal tip height and auricular height (0.01< EM>P /EM><0.05 ), but no differences on the head length, min frontal, interocular broad, nose length, mouth breadth, lip height, total height of head, and morphological facial height ( EM>P/EM> >0.05 ). The head breadth, intertragiar breadth, face breadth, extrinsic biocular breadth, physiognomic facial width and maximum circumference of the head extremely were significant difference ( EM> P/EM> <0.01 ). Most males and females were lissotrichous and black and the forehead hair was straight. Most of males and females were tint skin color and eye color were pitchy. In terms of eye slit direction，external angles were mostly higher and greater compared to the internal angles in direction of eyeslits, most of their palpebra superior had wrinkles, and eyelashes were short. Most males were convex type and most females were straight contour of nasal bridge, the nasal tip was upturned and nostril was orbicular-ovate. The upper lip was protruding and went beyond the lower lip. The total thickness of the lips were middling, the lobe types were mostly triangle. Most of the male heads were characterized by being hyperbrachycephaly and females were brachycephaly. The face forms were mainly featured by Euryprosopy and their noses were mesorrhine. Conclusion The traits on the head and face constitution between males and females of Miao Nationality in Xishui indicate existence of significant sex difference and the disparity. The males physical characters of the Miao in Liangcun township of Xishui city are most closely related to the Miao nationality of Wangka in Guizhou and the females is most closely related to the Bouyei nation Related Articles | Metrics

技术方法

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Combining ethylene diamine tetraacetic acid with N-butanol to prepare paraffin tissue sections from the rat with tibia bone cancer pain 2012, 43 (3):  430-432.  doi: 10.3969/j.issn.0529-1356.2012.03.026 Abstract ( )   Objective To explore a simple and effective method for preparing paraffin tissue sections using specimens of tibia bone cancer pain in rats. Methods Specimens of tibia bone were obtained from the rats of bone cancer pain model group and normal group. The specimens were decalcified with the heated ethylene diamine tetraacetic acid (EDTA), followed by the dehydration and clearing with N-butanol instead of traditional methods of decalcification in strong acid, dehydration through absolute alcohol and clearing in xylene. The specimens were stained with hematoxylin and eosin. Results The morphological structures of specimens in both groups were intactly preserved. Compared with the traditional method, the present method had the following advantages: the procedure was simple, the duration of dehydration and clearing was more flexible, and the paraffin blocks shown a better cutting performance. Conclusion It is a suitable and reliable method to jointly use EDTA and n-butanol in preparing paraffin tissue sections using specimens of tibial bone cancer pain in rats. Related Articles | Metrics