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2011, Volume 42 Issue 1
06 January 2011 Previous Issue    Next Issue
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Predegenerated common peroneal nerve regulates cells apoptosis/proliferation and their gene expressions in injured rat optic nerve 2011, 42 (1):  1-6.  doi: 10.3969/j.issn.0529-1356.2011.01.001 Abstract ( )   Objective To study the effect of pre-degenerated common peroneal nerve on apoptosis and proliferation in injured rat optic nerve. Methods One hundred and fifteen adult male rats were divided into normal, injured and transplantation group. Cell number, apoptosis/proliferation, proliferating cell nuclear antigen(PCNA) expression and apoptosis/proliferation related gene expression in rat optic nerves were examined respectively using HE staining, flow cytometry, Western blotting, gene chips and real-time PCR. Results Cells in the optic nerve of the injury group increased compared with that in the normal group, but was lower than that in the transplantation group. Compared with the normal group, apoptotic cells increased and proliferated cells decreased in the injury group; Compared with the injury group, apoptotic cells decreased and proliferated cells increased in the transplantation group. Compared with the normal group, PCNA expression was downregulated in the injury group, but PCNA expression was up-regulated in the transplantation group than that in the injury group. Compared with normal group, thirty seven apoptosis and proliferation related genes expressed differentially in the injury group; Compared with the injury group, three genes expressed differentially in the transplantation group. Conclusion The cellular apoptosis increases and the cellular proliferation decreases in the distal part of the optic nerve after the optic nerve injury. Autograft transplantation of the common peroneal nerve could reduce the cellular apoptosis and promote the cellular proliferation. In addition, some corresponding gene expressions changed. Related Articles | Metrics

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Expression of endothelin-B receptors in the microglia following cerebral ischemia in adult rats 2011, 42 (1):  7-13.  doi: 10.3969/j.issn.0529-1356.2011.01.002 Abstract ( )   Objective To explore the relationship between the endothelin-B(ET-B) receptors and cerebral ischemia by studying the expression of endothelin-B receptors in the activated microglial cells. Methods The rat model of middle cerebral artery occlusion (MCAO) was established by minimal invasive craniotomy. Eighty one adult male SD rats were randomly divided into ischemia group including 2 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days and 1 week after ischemia (at each time point, EM>n/EM>=9); a sham operation group (EM>n/EM>=9) and normal control group (EM>n/EM>=9). ET-B receptor immunoreactivity was observed in activated microglial cells following MCAO by double labeling with lectin. The expression of ET-B receptors mRNA and protein were investigated by Western blotting and real-time polymerase chain reactions (RT-PCR). Results ET-B immunofluorescence was markedly induced in the activated microglial cells in both infarcted and peri-infacted zones by double labeling with lectin at the 1st day, 2nd day, 3rd day and the 1st week, especially at the 3rd day and the 1st week after MCAO. In the control and sham operated rats, microglial cells appeared and relatively unactivated and showed a lack of ET-B immunofluoresecence. ET-B mRNA and protein levels were steadily upregulated from 2 hours to 12 hours (EM>P/EM><0.05) after MCAO as compared with the controls, peaking at the 6th hours. At the 1st week, ET-B mRNA and protein levels were only marginally elevated above the control level. Conclusion A major finding of this study is the massive accumulation of activated microglia and the induced expression of ET-B receptors in activated microglial cells following MCAO. It is suggested that ETs via ET-B may act on activated microglia and play an important role in cerebral ischemic injury bearing the receptor. Related Articles | Metrics

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Effect of astragalus injection on the expression of Caspase-3 after hypoxia/hypoglycemia and reoxygenation in hippocampal neurons of rats 2011, 42 (1):  14-21.  doi: 10.3969/j.issn.0529-1356.2011.01.003 Abstract ( )   Objective To investigate the effect of astragalus injection on the expression of Caspase-3 interrelated by apoptosis after hypoxia/hypoglycemia and reoxygenation in hippocampal neurons of rats. Methods Divided the hippocampal neurons cultured for eight days into four groups: normal control group(normal), hypoxia/hypoglycemia and reoxygenation group(model), astragalus injection solution group(solution) and astragalus injection group(astragalus). All groups were treated with reoxygenation after deprived of oxygen and glucose for 30 minutes except normal group. Immunohistochemistry was used to measure the number of Caspase-3 positive neurons, EM>in situ/EM> hubridization and RT-PCR were used respectively to measure the expression of Caspase-3 mRNA after hypoxia/ hypoglycemia and reoxygenation 0,0.5,2,6,24,48,72,120 hours. The experiment was repeated 6 times. Results The results of immunohistochemistry were similar to that in EM>in situ/EM> hybridization. It could be observed in normal control group that hippocampal neuronal nucleolus were integrated, cellular tuber was distinct and cytokinesis were dyed by yellow a lot; It could be observed in hypoxia/hypoglycemia and reoxygenation group that hippocampal neuronal nucleolus were crimple, cellular tuber was shrinked, large numbers of cytokinesis were dyed by yellow and there were yellow granules. Compared with normal control group, the numbers, the percentage and the average absorbance(AEM>A/EM>) of Caspase-3 protein and Caspase-3 mRNA positive neuronal cells every time points increased obviously in the hypoxia/ hypoglycemia and reoxygenation group except 0 hour and 0.5 hours(EM>P/EM><0.05), and peaked at 24 hours. The change of neuronal in configuration, numbers，percentage and the average absorbance of Caspase-3 mRNA positive neuronal cells in astragalus injection solution group were accorded with hypoxia/ hypoglycemia and reoxygenation group（EM>P/EM>>0.05）. Compared with hypoxia/hypoglycemia and reoxygenation group, the numbers, the percentage and the average absorbance of Caspase-3 mRNA were less obviously in the astragalus injection group except 0 hour and 05 hours(EM>P/EM><0.05). RT-PCR result showed that compared with normal control group, the average absorbance of expression of Caspase-3 mRNA in hippocampal neurons of rats every time points increased obviously in hypoxia/hypoglycemia and reoxygenation group except 0 hour and 0.5 hours (EM>P/EM><0.05), and peaked at the 24th hour; Compared with hypoxia/hypoglycemia and reoxygenation group, the average absorbance of expression of Caspase-3 mRNA every time points in astragalus injection solution group had no obvious change, however the average absorbance of expression of Caspase-3 mRNA in hippocampal neurons of rats every time points decreased obviously in the astragalus injection group except 0 hour and 0.5 hours(EM>P/EM><0.05). Conclusion Astragalus injection could inhibit the expression of Caspase-3 after hypoxia/hypoglycemia and reoxygenation, accord Related Articles | Metrics

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Differentiation of rat bone marrow mesenchymal stem cells into dopaminergic neurons after treatment with sonic hedgehog and fibroblast growth factor-8 2011, 42 (1):  22-26.  doi: 10.3969/j.issn.0529-1356.2011.01.004 Abstract ( )   Objective To explore the differentiation of rat bone marrow mesenchymal stem cells(BMSCs) into dopaminergic neurons EM>in vitro/EM> after induced by sonic hedgehog (SHH) and fibroblast growth factor-8 (FGF-8). Methods The BMSCs derived from rat bone marrow were cultured and passaged in vitro. After being induced by basic fibroblast growth factor(bFGF) for 24 hours, the BMSCs were incubated with SHH and FGF-8 for 12 days. Then the expressions of Nestin, neuron specific enolase(NSE), glia fibrillary acid protein(GFAP), tyrosine hydroxylase(TH), vesicular monoamine transporter-2(VMAT2), proliferating cell nuclear antigen(PCNA), Bcl-2, cyclindependent protein kinase2(CDK2) and Cyclin B1 were detected by immunocytochemical and immunofluorescence methods. Results Under the inverted microscope, the induced BMSCs showed typical neuronal morphological characteristics after induction, and they downregulated PCNA, CDK2 and Cyclin B1.The induced cells also expressed Nestin, NSE, TH and VMAT2 in high degree and GFAP in low degree. The percentages of induced cells and uninduced BMSCs positive for TH were (46 Related Articles | Metrics

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Expression of neurogenic differentiation factor increased after transient hypoxia of hippocampal neurons EM>in vitro/EM> 2011, 42 (1):  27-31.  doi: 10.3969/j.issn.0529-1356.2011.01.005 Abstract ( )   Objective To observe the influence of transient hypoxia on the expression of neurogenic differentiation factor(NeuroD) in cultured neurons and investigate its possible roles in neural regeneration. Methods Influence of transient hypoxia on the expression of NeuroD was analyzed on the outcome of embryonic rat neurons in culture. Cultures at five days were exposed to hypoxia. After different periods(3 hours, 6 hours) of hypoxia, RT-PCR was performed to examine the mRNA levels of NeuroD. Electron microscopy was performed to observe neuronal alterations. Dishes after 3 hours hypoxia were then returned to normal atmosphere for ensuing culture 96 hours. Immunohistochemistry was performed to examine cell proliferation by incorporation of 5-bromodeoxyuridine(BrdU). Results Following hypoxia for 3 hours in cultured neurons, NeuroD increased distinctly and the incorporation of BrdU revealed an accumulation of proliferating cells.Compared with the control group, NeuroD showed no much difference after hypoxia for 6 hours(EM>P/EM>>0.05). Neurons exposing hypoxia for 6 hours were damaged by electron microscope. Conclusion The expression of NeuroD increases post asphyxia in cultured neurons, following with cell proliferation. NeuroD seems to play a role in the process of neurogenesis. Related Articles | Metrics

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p38MAPK is involved in the cognition injury after sleep deprivation in rats 2011, 42 (1):  32-37.  doi: 10.3969/j.issn.0529-1356.2011.01.006 Abstract ( )   Objective To investigate the mechanisms of p38 Mitogenactivated protein kinase signaling pathway in cognition injury after sleep deprivation in rats. BR> Methods One hundred male SD rats were randomly divided into three groups: control group（EM>n/EM>=20），model group（EM>n/EM>=40）, inhibitor SB203580 treatment group（EM>n/EM>=40）. Sleep deprivation models were established by the modified multiple platform method. After sleep deprivation 1 day,3 days,5 days,7 days，morphological changes were observed by electron microscopy.The expressions of phosphorylated p38MAPK and interleukin 1β（IL-1β） proteins were detected with immunohistochemistry and Western blotting. The learning and memory functions were performed with Eightarm maze. Results Compared with control group,the morphous of nerve cells changed；the expressions of phosphorylated p38MAPK and IL-1β proteins increased；the learning and memory functions significantly decreased with time of sleep deprivation. Compared with model group, the morphological changes,t Related Articles | Metrics

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Influence of laminin in cell growth inhibition of curcumin on human hepatocellular carcinoma HepG-2 cells 2011, 42 (1):  38-44.  doi: 10.3969/j.issn.0529-1356.2011.01.007 Abstract ( )   Objective To study the influence of curcumin on cell growth and apoptosis in HepG-2 cell, in the presence of exogenous laminin(LN) to its receptor. Methods HepG-2 cells were cultured in DMEM containing 10% fetal bovine serum at 37℃ in a humidified atmosphere with 5% COSUB>2./SUB>The experiment was performed in four groups:the control group, LN group(20μg/L LN), curcumin group(40μmol/L curcumin),and combination group ( 20μg/L LN+40μmol/L curcumin ). Acid phosphatase assay(APA),flow cytometry（FCM） and Western blotting were used to detect the cell viability, apoptosis ratio, intracellular calcium concentration ,mitochondrial transmembrane potential,expression of proliferation-related proteinα-PKC and apoptosis-related protein poly ADP ribose polymerase(PARP), Caspase-3, p53 and Bcl-2 Results Compared with the control group ,LN group increased the viability rate. Curcumin(40μmol/L)and combination( 20μg/L LN+40μmol/L curcumin ) inhibited HepG-2 cells proliferation obviously, somewhat in a time dependent manner. After treatment with 40μmol/L curcumin and 20μg/L LN+40μmol/L curcumin for 48hours,the number of HepG-2 cells reduced, volums shrank, shape rounded and suspended mostly. The rates of late apoptosis and necrosis cells(%) were 97.04±1.50,98.02±1.35.Intracellular calcium concentration rised. Mean fluorescence intensity (MFI) of mitochondrial transmembrane potentials in HepG2 cells dropped. The expression of proliferation-related proteinα-PKC decreased. Curcumin could induce cleavage of PARP. The expression of apoptosis-related protein p53 increased, Caspase-3 decreased but Bcl-2 ha Related Articles | Metrics

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Selecting human fetal hepatic progenitor cells based on α-fetoprotein promoter activities 2011, 42 (1):  45-49.  doi: 10.3969/j.issn.0529-1356.2011.01.008 Abstract ( )   Objective To select hepatic progenitor cell clones from human fetal liver cells (FLCs) based on α-fetoprotein (AFP) promoter activities. Methods AFP promoter was amplified from human fetal liver genome by polymerase chain reaction (PCR) and cloned into pGL3-basic vector, forming the pGL3-AFP in a direct orientation for the promoters to drive firefly luciferase expression. To analyze the specificity of the AFP promoter, the pGL3-AFP and pRL-TK plasmids were co-transfected into HepG2, A549, and HeLa cells; the latter two cells do not express AFP. In order to obtain the homogenous cell populations, the proliferating colonies were isolated from clonal cultures of FLCs, and the AFP promoter activity of each clone was analyzed in each well of the cell lysates after co-transfection of pGL3AFP and pRL-TK plasmids for 48hours. Immunofluorescence was used to confirm the AFP expression in each clone. Results The results of PCR, restriction enzyme cutting and DNA sequencing confirmed that the pGL3-AFP had been constructed successfully. After co-transfection of the pGL3-AFP and pRL-TK plasmids, the AFP promoter activity was detected in HepG2 cells, but was weakly detected in A549 and HeLa cells, indicating that the AFP promoter could drive downstream firefly luciferase gene expression in AFP-expressing cells. Although the promoter activity in FLCs was lower than that in HepG2 cells, it was higher than that in AFP-non-expressing A549 and HeLa cells, further indicating that there were AFP-expressing cells among the FLCs. Five proliferating cell colonies were generated from clonal cultures of the FLCs, and the AFP promoter activity could be detected in one of the colonies. Immunofluorescence results revealed that the percentage of AFP positive cells was (99.1±0.6)% in this clone, indicating a hepatic progenitor cells clone. Conclusion The homogenous hepatic progenitor cell clones could be selected by AFP promot Related Articles | Metrics

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p73 activation is involved in regulation of Ara-C-induced apoptosis in human lung adenocarcinoma cells 2011, 42 (1):  50-53.  doi: 10.3969/j.issn.0529-1356.2011.01.009 Abstract ( )   Objective To study the apoptosis pathway of human lung adenocarcinoma cell line A549 induced by 1-β-D-arabinofuranosylcytosine (Ara-C) EM>in vitro/EM>. Methods A549 cells were incubated with Ara-C for 72hours EM>in vitro/EM>. Biological changes of apoptotic cells were studied by TUNEL staining. Morphological changes of the A549 cells treated with Ara-C were observed by transmission electron microscope. The expressions of p53 and p73 were investigated by Western blotting. Results 1.Apoptotic rates of A549 cells exposure to Ara-C studied by TUNEL staining were higher than that of the control (EM>P/EM><0.01). 2.Apoptosis body was apparently observed by transmission electron microscope. 3.Endogenous p73 but not p53 was induced and activated in dose-dependent manner upon Ara-C treatment by Western blotting. Conclusion Ara-C can effectively induce apoptosis of A549 cells. DNA damage-induced apoptosis of A549 cells treated by Ara-C is independent of functional p53-Up-regulation of p73 may play an important role that enhances the sensitivity of A549 cells to Ara-C and be p Related Articles | Metrics

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Effects of diethylstilbestrol on the expression of prolactin in mice lymphoid nodes 2011, 42 (1):  54-60.  doi: 10.3969/j.issn.0529-1356.2011.01.010 Abstract ( )   Objective To investigate the changes of prolactin (PRL) expression in lymphoid nodes of BALB/c mice treated with diethylstilbestrol (DES) in neonatal period and whether these changes differ from female mice to male ones. Methods Neonatal brood BALB/c mice whose weight discrepancies less than 5% were divided randomly into control group and DES group, and 25 pairs of female mice and 25 pairs of male mice were chosen in all. Within 24 hours after birth, mice in DES group were injected subcutaneously at cervix-backside with DES at intervals of 24 hours for 5 times, and mice in control group were injected with germ-free bean oil by the same way. Mice were killed on day 7, 21, 35, 49 and 63 after birth separately, and 5 pairs of female mice and 5 pairs of male mice were killed at each time point. The lymphoid nodes were taken out, imbedded with paraffin and sectioned serially in 5 μm. The expression of PRL in slices of lymphoid nodes was detected by immunohistochemistry technique. Results No matter in female or male mice, PRL positive cells were present in mice of both DES group and control group since day 7 until day 63 after birth. Prolactin positive cells were recognized as lymphocytes with typical morphological characteristics, and distributed in cortex and medulla. PRL expression in both groups increased in an age-dependent manner until day 35 after birth and then declined. At each the same time point, both the average absorbance (A EM>A/EM> ) and the integrate absorbance (I EM>A/EM> ) of PRL expression were higher in DES group than those in control group. In female mice, statistic differences were showed in A A on day 35 and in I A on day 21, 35 (EM>P/EM><0.001), 49 and 63 after birth between the two groups. In male mice statistic differences were showed in AEM> A/EM> on day 35 and in I EM>A/EM> on day 21 and 35 between the two groups. Conclusion In both female and male BALB/c mice PRL positive cells were present in their lymphoid nodes, and the level of PRL expression was the highest on day 35 after birth. Being reated with DES during neonatal period could lead an increased PRL expression in lymphoid nodes of BALB/c mice. The female BALB/c mice were much more sensitive to DES stimulation than male mice, and the effects of DES on female mice can continue to their adulthood at least. Related Articles | Metrics

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Cell cycle regulation of activated Akt in cardiomyocytes 2011, 42 (1):  61-64.  doi: 10.3969/j.issn.0529-1356.2011.01.011 Abstract ( )   Objective In order to study the effect of Akt on cardiomyocytes proliferation, the expression of Akt during postnatal development of rat heart was detected and the change of mitosis was examined after Akt was activated on cardiomyocytes. Methods The mRNA expression of Akt and the phosphorylation level of Akt protein during the postnatal developmental process (five cases per groups) were detected by RT-PCR and Western blotting analysis in rat hearts. H9c2(2-1) cardiomyocytes were originally isolated and cultured from rat BD1X heart.The Akt was activated by insulin stimulation and/or pCIS2-Akt-WT plasmid transfection. After that the expression of phosphor-histone H3(H3P) protein was detected by immunoprecipitation analysis on H9c2(2-1) cardiomyocytes. Results The mRNA expression of Akt and the phosphorylation level of Akt protein were down-regulated significantly（EM>P/EM><0.01）during postnatal 0-day-old group to 2-week-old group of rat heart. Actived Akt increased markedly the expression of H3P protein on H9c2(2-1) cardiomyocytes（EM>P/EM><0.01）. Conclusion Akt can promote mitosis of Related Articles | Metrics

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The compatibility study of osteoblast and the silicate-phosphate coating on the surface of the magnesium EM>in vitro/EM> 2011, 42 (1):  65-69.  doi: 10.3969/j.issn.0529-1356.2011.01.012 Abstract ( )   Objective To evaluate the growing condition of osteoblasts, which were cultured on the silicatephosphate coating on the surface of the magnesium and the biocompatibility between the coating and osteoblasts. Methods The osteoblasts were harvested by collagenase digesting method,and identified by immunofluorescence and Alizarin red staining method.Then the osteoblasts of the forth passage were inoculated on the material surface and the changes in form of osteoblasts on different surfaces were observed by scanning electron microscopy(SEM).At the same time the material leaching liquor was prepared to culture osteoblasts and the viabilities and differentiation of osteoblasts were detected by MTT and ALP method respectively. Results SEM showed that cells on the coating magnesium alloy growed well, and proliferated significantly,but cells were hardly finded on the naked group. The MTT and ALP analysis showed that 1%-5% of Si coating could promote the growth,proliferation and differentiation of osteoblast more obviously than other groups EM>in vitro/EM>. Conclusion The silicatephosphate coating on the surface of the magnesium had good compatibility with osteoblasts and was conducive to cell growth ,adhere, prolif Related Articles | Metrics

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Brain derived newotrophic factor gene-modified cortical neural stem cells of rat embryo and their effect on differentiation EM>in vitro/EM> 2011, 42 (1):  70-74.  doi: 10.3969/j.issn.0529-1356.2011.01.013 Abstract ( )   Objective To construct brain derived neurotrophic factor (BDNF) recombinant plasmid and transfect neural stem cells (NSCs), and observe its effect on differentiation of NSCs. Methods Rat cortical neural stem cells were cultured and identified for transfection. Recombinant plasmid with BDNF gene was constructed and identified by enzyme digestion and sequencing analysis. Non-liposome transfection pattern was used to transfect NSCs. Cells were divided into pcDNA3-1-BDNF group, pcDNA3-1 group and NSCs group without transfection. The expression of BDNF protein and mRNA were detected in infected NSCs by immunocytochemistry and RT-PCR. The infected NSCs were differentiated in medium containing 5% fetal calf serum, and identified by immunoreactive to special antibody, βIII-tubulin, glial fibrillary acidic protein (GFAP) and synaptophysin (SYP). The expression of SYP was observed during the differentiation of NSCs transfected with pcDNA3-1-BDNF. Results NSCs obtained in vitro were nestin positive. Recombinant plasmid with BDNF gene was successfully constructed. The expression of BDNF protein and mRNA of NSCs was significantly increased in pcDNA3-1BDNF group compared to the pcDNA3-1 group and NSCs group was found (EM>P/EM>0.05), but no significant difference between pcDNA3-1 group and NSCs group(EM>P/EM>＞0.05). The cells differentiated into βIII-tubulin positive and GFAP positive cells in all groups, to a lesser extent, also SYP positive in pcDNA3-1BDNF group at 4 days after differentiation. The proportion of βIIItubulin positive for neurons was significantly higher in pcDNA3-1-BDNF group than that in the pcDNA3-1 group and NSCs group at 7 days after differentiation (EM>P/EM>0.05), the number of SYP positive cells and the fluorescence intensity were increased in pcDNA3-1-BDNF group. Conclusion NSCs transfected with recombinant plasmid pcDNA3-1-BDNF have the capacity of producing BDNF, to promote direct Related Articles | Metrics

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Expression and clinical significance of EZH2 in hepatocellular carcinoma 2011, 42 (1):  75-79.  doi: 10.3969/j.issn.0529-1356.2011.01.014 Abstract ( )   Objective To study the expression of EZH2 gene in hepatocellular carcinoma (HCC) and its clinical roles. Methods Reverse transcription-polymerase chain reaction (RT-PCR)， immunohistochemistry, and Western blotting were used to detect the EZH2 mRNA and EZH2 protein expression in the tumor samples and surrounding non-cancerous liver samples，and then investigated the correlation between expression status and various clinicopathological parameters. Results The positive rate of EZH2 mRNA in HCC was 619%(26/42),the EZH2 mRNA positive rate in surrounding non-cancerous liver samples was 28.6% (12/42).EZH2 mRNA expression in HCC was significantly higher than that in surrounding noncancerous liver samples (EM>P/EM><0.01). Immunohistochemistry showed EZH2 protein expression in HCC was also significantly higher than that in surrounding non-cancerous liver samples(25/42 vs 9/42, EM>P/EM><0.01). Western blotting showed that EZH2 protein was expressed in all selected 18 HCC and surrounding noncancerous liver cases.The quantity of EZH2 protein in HCC was much more than that in surrounding non-cancerous tissue(0.90±0.30 vs 0.36±0.21，EM>P/EM><0.01).As the EZH2 and pathologic parameters were analysed, it was found that EZH2 mRNA transcript was elevated in poorly differentiated HCC compared with wellmoderately differentiated HCC,but there were no statistically significant correlations between EZH2 mRNA and tumor size, and intrahepatic/ extrahepatic metastasis. No significant difference was found between EZH2 protein and tumor size, differentiation, and intrahepatic/ extrahepatic metastasis.[WTHZ]Conclusion The exp Related Articles | Metrics

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Relationship of the expression of ZO-1 mRNA and protein with invasiveness, metastasis and prognosis in gastric carcinomas 2011, 42 (1):  80-85.  doi: 10.3969/j.issn.0529-1356.2011.01.015 Abstract ( )   P>Objective To investigate the significance of zonula occludens-1(ZO-1) mRNA and protein expression in invasion and metastasis and prognosis of human gastric carcinomas. Methods Real-time qPCR and SP immunohistochemistry on tissue microarray were used to detect 52 cases of ZO-1 mRNA and 228 cases of protein expression in gastric cancer. Results In contrast to normal epithelium, 81.1% gastric carcinoma tissues showed different degrees of aberrant expression of ZO-1 from membrane translocated to cytoplasm. The ZO-1 aberrant degree in the mucosal layer of gastric carcinoma cells in the case of well-differentiated, invasiveness depth confined to muscularis, without lymph node metastasis and early gastric cancer was lower than those in poorly differentiated, invasiveness depth breakthrough to muscularis, with lymph node metastasis and progressed gastric cancer, respectively (EM>P/EM>0.05). The ZO-1 aberrant degree in the invasive front area of gastric adenocarcinoma cell was higher than that in mucosa, but in lymph node metastases lower than that in invasive front area (P0.05). Postoperative survival rate in case with ZO-1 aberrant expression in mucosal layer of gastric carcinoma was lower than that in ZO-1 normal expression(EM>P/EM>0.05). However, ZO-1 aSUP>+/SUP> mRNA expression between gastric adenocarcinomas in mucosal layer and normal gastric epithelial cells in their corresponding adjacent normal gastric mucosa was not significantly different. ZO-1 a+ mRNA expression in mucosal layer of gastric adenocarcinomas was not significantly correlated with age, sex, histological type, the invasive depth and status of lymph node metastasis. Conclusion ZO-1 aberrant expression involved in the progress of gastric adenocarcinoma, but ZO-1 mRNA expression was not significantly different. These results suggested that hot the change of ZO-1 mRNA quantity but the ZO-1 aberrant expression associated withcarcinog Related Articles | Metrics

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New discovery of a malignent cell line with telocentric chromosomes derived from nude mouse transplanted by ovarian epithelial carcinoma 2011, 42 (1):  86-90.  doi: 10.3969/j.issn.0529-1356.2011.01.016 Abstract ( )   Objective According to the usual method to establish tumor cell line, tissue of human ovarian epithelial carcinoma was transplanted to the immunodeficient mice and a immortalized cell line was established by primary culture, so to provide one more experimental model for the investigation of human carcinoma. Methods The cancer tissue from a patient with ovarian papillary serous carcinoma IIIcG3 was transplanted subcutaneously into node mice. A suspension cell line was established by primary culture EM>in vitro/EM>. The morphology of the cell line was observed by light microscope and electro microscope. Its biologic characteristics were investigated by growth curve, karyotype analysis, soft agar culture and nude mice transplantation. Results This cell line was named WSZ.It has been maintained for over one hundred passages. It was in suspension condition with vigorous growth.The population doubling time of cells in logarithmic growth phage was about 15.8 hours. The number of chromosomes was from 16 to 135 and 68 or 69 was multiple. Karyotype analysis showed that almost all the chromosomes were telocentric. It had a clone forming rate of 89.3% and could form clones in soft agar.It was demonstrated that toumor could be formed under the skin of nude mouse with cell number of 10SUP>6/SUP> while in non-obese diabetes（NOD) serve combined immuno-deficiency(SCID) only 100 cells were needed. Conclusion A malignet cell line with telocentric chromosomes derived from nude mouse transplanted by ovarian epithelial curcinoma was established, it is named WSZ is line, that possesses typical malignent characteristics. WSZ had stable infinite proliferation ability. WSZ is a cell line with high malignant. Related Articles | Metrics

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Effects of melamine on histological structure and activities of superoxide dismutase，catalase and content of maleic dialdehyde in kidney of filial mice 2011, 42 (1):  91-98.  doi: 10.3969/j.issn.0529-1356.2011.01.017 Abstract ( )   Objective To investigate the changes of kidney weight and histological structure，activities of superoxide dismutase(SOD)，catalase(CAT) and content of maleic dialdehyde(MDA) in kidney after the developing filial mice received intragastric administration of melamine, and to evaluate the effects of melamine on kidney development of filial mice．Methods Totally 160 filial mice at the seventh day respectively received 0.3，0.8 and 1.3g/(kgI>&#/I>8226;d) doses of melamine in the three experimental groups by intragastric administration，while the control group were given the same amount of saline．The samples were harvested at day 6，12，18，24 and 30 respectively． The shape and weight of kidneys were determinded，and the activities of SOD，CAT as well as content of MDA were detected by colorimetry．In addition，the changes of the renal lesions were observerd by optical microscope in all groups．Results At day 18-30 after the administration of melamine， kidneys of some experimental filial mice were swelling with reddish brown appearance，and the body weights were significantly higher than those of the control group．In 0.3g/(kgI>&#/I>8226;d) and 0.8g/(kgI>&#/I>8226;d) group，SOD and CAT activities in kidney increased and were higher than that of the control group in the early time，then decreased and were significantly lower than that of the control group after 24 days， while MDA content in kidney decreased and lower than that of the control group in the early time，then increased and much higher than that of the control group after 24 days. In 1.3g/(kgI>&#/I>8226;d) group, SOD and CAT activities in kidney were always significantly lower than that of the control group，and MDA content was always much higher than that of the control group．With the extension of administration days，cortex and medulla of kidney of experimental filial mice appeared with some difference．The epithelial cells of renal tubule and collecting duct were swelling，vacuolization and necrosis；expansion and hemorrhage of capillary；colloid-like material appeared Related Articles | Metrics

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SCF/c-KIT signal regulates the proliferation and division of alpaca (EM>lama pacos/EM>) hair follicle melanocytes and its mechanism 2011, 42 (1):  99-103.  doi: 10.3969/j.issn.0529-1356.2011.01.018 Abstract ( )   Objective To research the cytological mechanism of the genesis of different coat color in alpaca by investigating the regulation mechanism of the SCF/c-KIT signal on the cell division, proliferation and distribution of alpaca hair follicle melanocytes. Methods We selected eight mature female alpacas, four with pigmented coat color and four with natural white coat color. Immunohistochemistry was used to identify the expression distribution of SCF and c-KIT receptor. Comparative quantitative RT-PCR was used to determine expression levels of SCF and c-KIT genes. Results The expression of SCF and c-KIT immunoactivity was detected in different coat color skin, but the expression level and localization were different in the hair follicle. ΔΔEM>Ct/EM> method was used for quantitative analysis of the expression levels of SCF and c-KIT genes in different coat colors of alpaca skin. The comparative expression of gene SCF in colored skins was 2.41 times of white skins, and the comparative expression of gene c-KIT in colored skins was 1.20 times of white skins. Conclusion SCF/c-KIT signal regulates the hair follicle melanocytes proliferation and division. The mature degree and mounts of melanocytes in hair follicle play central role in the genesis of coat color of alpaca. Furthermore, the differential distribution of melanocytes in hair follicle is mainly regulated by the Related Articles | Metrics

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Effects of sericine on growth hormone/insulin-like growth factor-1 axis of testis in type 2 diabetes mellitus rats 2011, 42 (1):  104-109.  doi: 10.3969/j.issn.0529-1356.2011.01.019 Abstract ( )   Objective To study the effects of sericine on growth hormone(GH)/insulin-like growth factor-1(IGF-1) axis of testis in type 2 diabetes mellitus rats model. Methods Forty male SD rats were randomly divided into 4 groups(EM>n/EM>=10): normal control group, diabetes model group, sericine treatment group and positive control group. The type 2 diabetes model rats were induced by continuous intraperitoneal injection of streptozotocin and blood glucose≥16.7mmol/L was taken as the criterion one week after injection. After successfully establishing the diabetes animal models, the rats in diabetes model group were not given any more treatment, the rats in sericine treatment group were lavaged with sericine ［2.4g/(kgI>&#/I>8226;d), 35d］, the rats in positive control group were lavaged with metformin ［55.33mg/(kg[DK]I>&#/I>8226;d), 35d］. ELISA was used to detect the testosterone, GH and IGF-1 level in serum; Immunohistochemical staining, Western blotting and RT-PCR were respectively used to detect the GH, growth hormone receptor (GHR) and IGF-1 expression in testis. Results Sericine could significantly decrease GH level in serum and down regulate GH expression in testis; remarkably increase testosterone and IGF-1 level in serum, up regulate GHR and IGF-1 expression in testis (EM>P/EM>0.05, EM>P/EM>0.01). Moreover, there had no obvious differences between sericine treatment group and positive control group (P＞0.05). Conclusions Sericine can protect and improve reproductive disfunction of diabetes rats by regulati Related Articles | Metrics

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Development of CD-positive cells and CD20-positive cells in human fetal palatine tonsil 2011, 42 (1):  110-113.  doi: 10.3969/j.issn.0529-1356.2011.01.020 Abstract ( )   Objective To study the development of T and B lymphocytes in human fetal palatine tonsil. Methods Totally 22 samples of human fetal palatine tonsil from the 9th to 20th gestational week (GW) were collected. T and B lymphocytes were displayed with cluster of differentiation（CD)，anti-CD3 antibody and anti-CD20 antibody by immunohistochemical ultrasensitive-SP method. The immunoreactive positivecells were counted with BioMiaspro photographs analysis software and analysised with statistics software.Results In the 9th GW, there were rare CD3-positive cells and CD20-positive cells, distributed in the mesenchyme under the mucous membrane epithelium. In the 14th GW, the amounts of CD20-positive cells were significantly increasing, part of them were converged in primary lymphoid nodule,others were located in the diffused lymphoid tissue and crypt epithelium. At the same time, the amounts of CD3-positive cells were still to remain less, but a few of them were also infiltrated into the crypt epithelium.In the 17th GW, the number of primary lymphoid nodule was increaseing. In the 20th GW, the quantities of CD3-positive cells were significantly increasing, which were diffused distribution and formed grouping in the lymphoid tissue of lamina propria. The two types of positive-cells were increasing gradually with fetal age growing（EM>P/EM><0.05 or EM>P/EM><0.01）,but the numbers of CD20-positive cells were majority (EM>P/EM><0.05). Conclusion In the 9th GW, T and B cells have emerged in the human fetal palatine tonsil; In the 14th GW, B cells converged and Related Articles | Metrics

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Distribution and morphological characteristics of testis lobule micrangium in adult Tibetan plateau sheep 2011, 42 (1):  114-118.  doi: 10.3969/j.issn.0529-1356.2011.01.021 Abstract ( )   Objective To study the architecture of the testis lobule micrangium and its functional relationship in 30 adult Tibetan plateau sheep. Methods The ABS vascular proplasm, ink-gelatinum paraffin section and replica scanning electron microscopic methods were used.[WTHZ]Results The small arteries from the centripetal or centrifugal artery walked through the polygonal tissue space，and so as the distribution of the small veins. The interstitial capillary also went along the longitudinal wall of the seminiferous tubule and formed ladder-like capillary network, and give branches interconnected each other to form peritubular capillaries. The peritubular capillaries also formed interstitial capillary plexus around the Leydig cells; Moreover, the broad and different level communications were found in microvasculature around the seminiferous tubules of the testis lobule. There were obvious imprints of the smooth muscles on the surface of the casts of the intralobular small arteries, the interstitial arteriole and the precapillary arteriole. On the surface of the casts of the interstitial arteriole and the precapillary arteriole, there were obvious imprints of the endothelial nuclei as well.[WTHZ]Conclusion Several testis lobule micrangium structural features are abserved in Tibetan plateau sheep. It could be the result of their living in lower-oxygen on Tibetan plateau from generation to generation. Related Articles | Metrics

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Induction of connective tissue on the differentiation of fetal bone mesenchymal stem cells into epithelial cells 2011, 42 (1):  119-123.  doi: 10.3969/j.issn.0529-1356.2011.01.022 Abstract ( )   Objective To observe the mechanism of the bone marrow mesenchymal stem cells (BMSCs) induced by connective tissue differentiate into epithelial cells. Methods Content of epidermal growth factor (EGF) in amnion removed epithelium and foetus intestinal connective tissue were detected by ELISA.BMSCs obtained from fetus aged 16-20 weeks were isolated, cultured and proliferated EM>in vitro/EM>. The DAPI marked P3-BMSCs were planted on the amnion, and added supernatant of intestinal connective tissue or amnion supernatant( both contained final concentration of 30ng/ L EGF)， and exogenous EGF 30μg/L， 30 ng/L， 0， group 1.5 respectively. At the cultivated 7th day, the expressions of cytokeratin(CK)、CK20 in BMSCs were observed with immunofluorescence method and CLSM. Results There were(320.22±0.257)pg，(299.20±0.994)pg EGF in the fetal intestinal connective tissue or amnion/ 100mg respectively. After induced by amnion and supernate of intestinal connective tissue or amnion, the CK and CK20 positive cells in BMSCs were obviously stronger than that in groups induced by amnion and exogenous EGF and in group induced only by amnion, EM>P/EM>0.01 The CK20 positive cells in group induced by amnion and supernate of intestinal connective tissue were much more high than that in group induced by amnion and supernate of amnion,EM> P/EM>0.05 The difference of CK and CK20 positive cells was not statistically significant among the group induced by amnion and exogenous EGF and in group induced only by amnion. Conclusion The direct contact may be one of mechanism tnat amnion induces BMSCs to differentiate int Related Articles | Metrics

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Influence of experimental cryptorchidism on Leydig cells in adult rats 2011, 42 (1):  124-0.  doi: 10.3969/j.issn.0529-1356.2011.01.023 Abstract ( )   Objective To investigate the change of the number, morphology and function of Leydig cells after induction of cryptorchidism. [WTHZ]Method Adult rats were induced cryptorchidism by confining the testes of both sides into the abdominal cavity. They were decapitated at intervals of 3, 7, 20 and 28 days postoperation (6 rats in each group). Sham group as the control. The number and morphological changes of Leydig cells were detected by the method of stereology. Application of immunohistochemistry detected of the expression of cell division cycle 25 A （CDC25A）, stem cell factor （SCF）, androgen receptor （AR）, estrogen receptor α（ERα)in Leydig cells. [WTHZ]Result The volume fraction of testicular interstitium increased significantly, but the absolute volume of testicular interstitium unchanged after induction of cryptorchidism. The number and morphology of Leydig cells did not change significantly. Immunohistochemical staining showed CDC25A, SCF strongly expressed in the area of Leydig cells, and the cryptorchidism had no significant effect on their expression. AR had weak staining in the cytoplasm of Leydig cells, and the intensity of AR expression no significant changes between different groups. The expression of ERα in Leydig cells in the control group was weak, almost no positive nuclei. But the percentage of Leydig cells which has ERα positive nucleus increased significantly with the prolongation of cryptorchidism time. It had reached 93.6% after 28 days cryptorchidism. [WTHZ]Conclusion Experimental cryptorchidism has no s Related Articles | Metrics

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Antifertility effect in mice after vaccination with plasmid DNA encoding mouse sperm equatorial segment protein 2011, 42 (1):  129-132.  doi: 10.3969/j.issn.0529-1356.2011.01.024 Abstract ( )   Objective To determine the feasibility of pcDNA3-1/mESP as a potential immunocontraceptive antigen. Methods Eukaryotic expression plasmid pcDNA3-1/mESP, encoding N-terminal of mouse sperm equatorial segment protein (mESP), was constructed. Two groups of mice（EM>n/EM>=8/group） were inoculated with plasmids of pcDNA3-1/mESP and pcDNA3-1 respectively. RT-PCR， ELISA and double immunofluorescence assay were performed to observe pcDNA3-1/mESP expression and immune response in the inoculated mice. Mouse mating test was employed to study fertility of the experimental and control group. Results mESP cDNA positive band was detected in muscle from mice immunized with pcDNA3-1/mESP，which identified the expression of the recombinant plasmid. The pcDNA3-1/mESP induced specific antibody against mESP in the experimental mice. Significant difference of mean litter size between experimental and control groups was observed, while the rate of fertility did not decrease. Conclusion This study indicates the need to strengthen DNA vaccines in order to obtain Related Articles | Metrics

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Database establishment of children head sample slices 2011, 42 (1):  133-136.  doi: 10.3969/j.issn.0529-1356.2011.01.025 Abstract ( )   Objective To establish the image database of the digital virtual chinese children craniofacial structure anterior to growth peak time with 0lmm intervals. Methods An 8-year-old anterior to growth peak time, middle growth and no deformity boy skull was selected to establish the visible chinese boy craniofacial structure image database. After 30 days of embedding, fixing and freezing, the slice-cutting of the skull was performed using a machine in -13℃ homothermic cryogenic laboratory in Research Center for Sectional and Imaging Anatomy, Shandong University School of Medicine. The milling was perfomed with 0.1mm at intervals. The Canon EOS-1DS MarkII camera was used to produce all the images. Results All original image data were saved as RAW format, which was transferred to PSD and JPG format for sharing and commumication more generally. The digital virtual chinese children craniofacial structure dataset gained a total of 2 150 images. An image size with RAW format was approximate of 20.4MB and the whole craniofacial structure images researched 4287GB. We saved 3 versions for variously oriented studies about boy craniofacial structure. Conclusion We established the serial anatomical cross-sectional image data of the craniofacial structure of the children anterior to growth peak time in succeeded. Compared to other available data, the dataset with at intervals 0.1mm were demonstrated to be of more efficient anatomical informa Related Articles | Metrics

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Anatomical study of basilar artery EM>in vivo/EM> 2011, 42 (1):  137-140.  doi: 10.3969/j.issn.0529-1356.2011.01.026 Abstract ( )   Objective To observe and measure the basilar artery EM>in vivo/EM> by 3.0 T magnetic resonance angiography(MRA). Method A total of 108 subjects experienced MRA and conventional MRI examination, the sex ratio was 58 male to 50 female, and the age ranged from 4 to 63 years ［mean (39.86±14.57) years］. Observing and recording the intracranial vertebrobasilar artery morphological type, measuring the length and lumen diameter of the basilar artery. Statistical testing was performed. Results The common types of basilar artery followed by typeⅠⅠ, typeⅡRⅢ, typeⅡRⅠor typeⅡLⅠ. The average straight line length of the basilar artery was (2.35 ± 0.30)cm. The straight line length of the basilar artery in men was longer than that in women (EM>t/EM>=4.19,EM> P/EM><0.000 1). Straight line length of the basilar artery was correlated with age (P <0.05, r=0.28). The number of the bending basilar artery was 51, in addition to 1 case of bilateral bending, the others were unilateral. The average diameter of the basilar artery was (0.26 ± 0.05)cm. there was no difference between the diameter of upper, middle and lower segment of basilar artery (EM>χ/EM>SUP>2/SUP> = 3.13, EM>P/EM>>0.05). The average diameter of th Related Articles | Metrics

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Analysis of cerebral glucose metabolism in mouse of Alzheimer’s disease model by Micro-PET 2011, 42 (1):  141-143.  doi: 10.3969/j.issn.0529-1356.2011.01.027 Abstract ( )   Objective To evaluate the cerebral glucose metabolism of Alzheimer’s disease(AD) model mouse by SUP>18/SUP>F-FDG and MicroPET，and develop an imaging technique to evaluate AD drugs therapeutic effect on animal models. Methods The 9 mice of control group, APPSUP>swe/SUP>/PSΔE9 doubletransgenic group and aricept treatment group［2 mg/(kgI>&#/I>8226;d)］ were injected SUP>18/SUP>F-FDG by peritoneal cavity and scanned by Micro-PET. The glucose uptake value of cerebrum，frontal cortex and temporal cortex region of interest(ROI) were analysed by IRW software. Results The images of SUP>18/SUP>F-FDG and Micro-PET could be used to detect the cerebral glucose metabolism of AD mouse model. We found that the cerebral glucose metabolism was decreased in the APPSUP>swe/SUP>/PSΔE9 doubletransgenic mice, which was consistent with phenotype of the human AD. Aricept treatment improved the cerebral glucose metabolism of AD disease model mice. Conclusion The imaging analysis using SUP>18/SUP>F-FDG an Related Articles | Metrics