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2016, Volume 47 Issue 6
06 December 2016 Previous Issue    Next Issue

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Triptolide promoting colon cancer CT26 cell death through induction of autophagy ZHANG Tian-jiao HAN Sen ZHANG Wei ZHAO Fu-jian LI Jian* 2016, 47 (6):  774-778.  doi: 10.16098/j.issn.0529-1356.2016.06.009 Abstract ( )   Objective To investigate the relationship of autophagy and cell death in CT26, a cell line from mouse colon cancer was used and treated by triptolide. Methods Immunofluorescence staining, LC3 adenovirus vector (mRFP-GFP-LC3), and Western blotting methods were adopted to examine autophagy flux in CT26 cells. The AQueous one solution cell proliferation assay(MTS) and AnnexinV-FITC/PI reagents were used to evaluate the cells inhibitions rate, apoptosis and cell death rate of CT26 cells after treated by triptolide, rapamycin, and triptolide combined with rapamycin. Results The CT26 cells showed significant autophagy flux induced by triptolide and activating cell autophagy improved the late CT26 cell apoptosis rate. Compared with the group of triptolide, the group of triptolide combined with rapamycin obviously increased CT26 cell death. Conclusion Triptolide promotes colon cancer CT26 cell death through induction of autophagy. References | Related Articles | Metrics

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Anti-tumor effects of chloroquine on S180 tumor-bearing mice and its mechanisms YUE Li-ling* MEI Qing-bu ZHU Wen-bin ZHANG Wei LIU De-shui LIU Ying 2016, 47 (6):  779-784.  doi: 10.16098/j.issn.0529-1356.2016.06.010 Abstract ( )   Objective To investigate the anti-tumor function of chloroquine on bearing cancer mice S180 and its possible mechanism. Methods Forty S180 tumor-bearing mice were divided into chlorquine, low dose group, middle dose and high dose group. The inhibiting effect of different concentration chloroquine on sarcoma 180 was observed with in vivo. The morphological changes in tumor cells of tumorbearing mice induced by chloroquine was observed under the transmission electron microscope. The apoptosis of tumor cells induced by chloroquine were tested with flow cytometry. The expression of apoptosisrelated factors Bcl-2, cytochrome-C and Cle-Caspase-3 was detected by immunohistochemistry. Western blotting was used to detect the protein levels of Bcl-2, Cle-Caspase-3 in tissue and the changes of Cytochrome-C in mitochondria and cytoplasm. Results Compared with the control group, the growth rate, the xenon graft volume and weight decreased obviously in chloroquine group (P<0.05). Apoptotic changes and apoptotsis body were discovered obviously in chloroquine group under the electron microscope. Chloroquine induced the apoptosis of bearing cancer mice S180 cell in every group, down-regulated the expression of anti-apoptotic factors Bcl-2 and up-regulated the expression of apoptotic genes Cle-Caspase-3 (P<0.05). Chloroquine reduced the mitochondria Cytochrome-C protein levels and increased the cytoplasm cytochrome-C protein levels (P<0.05). Chloroquine promoted the release of cytochrome-C to cytoplasm. Conclusion Chloroquine has the anti-tumor effect on the growth of bearing cancer mice S180 cell. It may affect the mitochondria apoptosis pathway and thus play a role in anti-tumor. References | Related Articles | Metrics

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Expression and significance of hypoxia inducible factor 2α together with CITED2 and PTPRZ1 in thyroid papillary carcinoma SUN Lin GAO Bin WANG Dong-mei ZHANG Wen-jing ZHANG Jing WU Jing-fang* 2016, 47 (6):  785-790.  doi: 10.16098/j.issn.0529-1356.2016.06.011 Abstract ( )   Objective To analyze the role and clinical significance of hypoxia inducible factor 2α(HIF-2α),CITED2 and PTPRZ1 during the development of papillary thyroid carcinoma (PTC) by detecting the expressions of HIF-2α,CITED2 and PTPRZ1 in the PTC tissues. Methods The expressions of HIF-2α,CITED2 and PTPRZ1 proteins in 50 cases of PTC were detected by Western blotting and immunohistochemical SP method, and the relationships with the clinical pathology characteristics were analyzed.The transcriptional level changes of HIF-2αmRNA,CITED2mRNA and PTPRZ1mRNA were tested by real-time reverse transcription polymerase chain reaction. Results The positive expression rate of HIF-2α CITED2 and PTPRZ1 in PTC tissues were obviously higher than that in thyrvid adenoma(TA) group and normal thyroid(N) group（P＜0.01）and expressions between N and TA were no significant difference （P＞0.05）. The expressions of HIF-2α, CITED2 and PTPRZ1 were unrelated to the age, sex and size of the tumor (P>0.05) of PTC patients, but associated with the lymph node metastasis and pathological stage (P<0.05). The transcriptional expression levels of the HIF-2α mRNA, CITED2 mRNA and PTPRZ1 mRNA in PTC tissues were significantly higher than in TA and normal tissues adjacent to the tumors with statistical significance(P<0.05), but there was no statistical difference between in TA and normal tissues adjacent to the tumors (P>0.05). Conclusion The higher expressions of HIF-2α, CITED2 and PTPRZ1 in PTC have a positive correlation with the lymph node metastasis, size and clinicopathologic stage. HIF-2α may promote the occurrence, development, invasion and metastasis of PTC by the higher expression of CITED2 and PTPRZ1.All of them play an important role in the process of PTC, include the occurrence, development, invasion and metastasis. References | Related Articles | Metrics

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Molecular mechanism of cell cycle arrest induced by Notch3 in human small cell lung cancer H446 cells LIU Hui CHEN Hong-lian YUAN Lei* 2016, 47 (6):  791-795.  doi: 10.16098/j.issn.0529-1356.2016.06.012 Abstract ( )   Objective To investigate the effect of Notch3 on proliferation and the cell cycle in human small cell lung cancer H446 cells and its possible mechanism. Methods H446 cells were stably transfected with human Notch3 and HES1 expressing plasmid separately. Short hairpin RNA (shRNA) was used to knock down MDM2 in H446 cells. The cell cycle was studied by flow cytometry. The mRNA expression levels of Notch3 and MDM2 were measured by RT-PCR. Western blotting was performed to determine the protein expression levels of Notch3, hairy and enhancer of split 1(HES1), murine double minute 2(MDM2), Rb and P21. Results Notch3 suppressed proliferation and induced cell cycle G0/G1 arrest in H446 cells(P<0.05), and up-regulated HES1 and down-regulated MDM2 at protein levels(P<0.05). High expression of HES1 resulted in a lower protein level of MDM2(P<0.05) without affecting the mRNA expression of MDM2. MDM2 knockdown suppressed proliferation and up-regulated Rb and P21 at protein levels(P<0.05).  Conclusion Notch3 may suppress proliferation and induce cell cycle arrest by up-regulating HES1, Rb and P21, and down-regulating MDM2. References | Related Articles | Metrics

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Distribution of cardiac telocytes in the aged rat heart XU Sha-sha LIAO Zhao-fu CAI Dong-qing* 2016, 47 (6):  802-806.  doi: 10.16098/j.issn.0529-1356.2016.06.014 Abstract ( )   Objective To observe the morphology and distribution of cardiac Telocytes (CTs) in the aged rat heart and the difference compared with young rat heart. Methods The transmission electron microscopy combining bio-image analysis and immunofluorescence staining were applied to analyze the morphology and distribution of CTs in the atrium, medium and apex part of 24-month-old rat heart semiquantitatively. Results A lot of cells with unique morphology of CTs were found in interstitial tissue of cardiac myoctyes and microvessels in the atrium, medium and apex part. The morphology parameters of CTs were similar in the three representative dimensions of heart. The difference is not statistic significance. The immunofluorence staining documented that aged CTs as well as young CTs expressed c-Kit and CD34. In aged heart, the CT density in the atrium part（47.65±4.01 cells/mm2）was significantly higher than that of the apex part (33.68±2.53 cells/mm2) and the medium part(26.49±2.11 cells/mm2) (P<0.05). In addition, it was found that the distributional pattern of CTs in the three representative dimensions of aged heart was similar to that of young heart. However, the CT density in the three representative dimensions of aged heart is significantly higher than that of the parallel areas of young heart (P<0.05). Conclusion CTs are found in the aged rat heart. The CTs density of the atrium part is significantly higher than that of the apex part and the medium part in aged heart. In addition, the distributional pattern of CTs in aged heart is similar to young heart. However, the CTs density of aged heart in atrium, medium and apex part is significantly larger than that of parallel areas of young heart respectively. References | Related Articles | Metrics

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Regulatory effects of sericin on pancreatic insulin PI3K/Akt signaling pathway in type 2 diabetic rats LIU Mei-xiao SUN Yi-chan HOU Li-na YANG Song-he CHENG Lu-yang SONG Cheng-jun* CHEN Zhi-hong* 2016, 47 (6):  807-811.  doi: 10.16098/j.issn.0529-1356.2016.06.015 Abstract ( )   Objective To investigate the effects of sericin on the blood glucose concentration by regulating pancreatic insulin P13K/Akt signaling pathway. Methods Thirty-six male SD rats were randomly divided into normal control group, diabetes mellitus model group and sericin treatment group, with 12 rats in each group. The type 2 diabetes mellitus rats model was induced by high calorie and high sugar diet plus intraperitoneal injection of streptozotocin (35mg/kg, 2 times, 1 time a day), and the standard for successfully establishing type 2 diabetes model was fasting blood glucose≥11.1mmol/L. The model rats were treated with sericin at the dose of 2.4g/kg/d for 35days by gastrogavage method. ELISA was used to detect the adiponectin level in serum, Western blotting and real time fluorescence quantitative PCR to detect the expression of insulin receptor (IR), insulin receptor substrate-1 (IRS-1), phosphatidylinositol 3-kinase (PI3K) and protein kinase B (Akt/PKB) protein and mRNA in pancreatic tissues. Results Compared with model group, the serum adiponectin level, IR, IRS-1, PI3K and Akt/PKB protein and mRNA expression in pancreatic tissues of rats in sericin treatment group increased markedly (P＜0.01,P＜0.05). Conclusion Sericin can improve the abnormal changes of PI3K/Akt signaling pathway by up regulating IR, IRS-1, PI3K and Akt expression in pancreas of diabetes mellitus rats; Thereby, sericin play a role in lowering blood glucose. References | Related Articles | Metrics

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Expression patterns of islet-1 in developing mouse embryonic hearts CAI Yu-jin JING Ya* SONG Li YANG Yan-ping CUI Hui-lin LI Ai-hong GUO Rui 2016, 47 (6):  812-817.  doi: 10.16098/j.issn.0529-1356.2016.06.016 Abstract ( )   Objective To observe the expression patterns of islet-1(ISL1) in developing mouse embryonic hearts and the morphogenesis of the heart, the second heart field and foregut endoderm. Methods Serial sections of eighteen mouse embryonic hearts from embryonic day(ED) 8 to ED 13 were stained immunofluorescently or immunohistochemically with antibodies against myosin heavy chain(MHC), ISL1, proliferating cell nuclear antigen(PCNA) and α-smooth muscle actin(α-SMA). Expression of ISL1 was measured with Western blotting in mouse embryonic hearts from ED11 to ED14. Results At ED9, ISL1-expressing cardiac progenitors was detected at the distal wall of the outflow tract (OFT). At ED 10, ISL1 positive cells extended into the proximal wall of OFT from the distal and distributed in the myocardium of venous sinus. From ED11 to ED12, ISL1-expression reached the highest level in embryonic hearts. ISL1 positive cells were mainly distributed at the sinus node and the venous valves on venous pole of the heart but were detected at the distal wall of OFT, the developing intrapericardial aorta, pulmonary trunk and aortic-pulmonary septum on the arterial pole of the heart. At the same time, the length of solid corn stretching out from foregut endoderm reached the longest and was surrounded by ISL1-expressing progenitors of anterior component of the second heart field (aSHF). The density of ISL1 positive cells in aSHF was significantly higher than that of posterior component of the SHF. At ED13, ISL1-expressing progenitors were dramatically reduced both in the heart and the SHF, while the solid endoderm cord was gradually eliminated. Conclusion ISL1 positive progenitor cells are mainly detected from ED9 to ED13 in mouse embryonic hearts. The extent of ISL1 positive cells distribution in mouse embryonic hears is accompanied by the morphogenesis of the SHF and foregut endoderm. References | Related Articles | Metrics

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Development of pulmonary endoderm-associated second heart field in mouse embryo SHI Liang LI Hui-chao CHEN Hao YANG Yan-ping JING Ya* 2016, 47 (6):  818-823.  doi: 10.16098/j.issn.0529-1356.2016.06.017 Abstract ( )   Objective To explore the morphological relationship and mechanism of pulmonary endoderm with the development of the prepharyngeal mesenchyme from second heart field during the outflow tract septation in mouse embryonic heart. Methods Both the immunohistochemical and immunofluorescence staining methods were used to observe forkhead box A2(Foxa2), islet-1(ISL-1), patched-1(Ptc1), patched-2(Ptc2), α-smooth muscle actin(α-SMA）and myosin heavy chain(MHC) distribution in serial sections of six mouse embryos each day from embryonic day(ED) 9 to embryonic day(ED) 13. Results At ED 9-ED 9.5, locally thickened ISL-1 positive endoderm in the ventral foregut wall predicted the initiation of pulmonary endoderm differentiation. As soon as the lanryngo-tracheal groove from pulmonary endoderm was initiated, ISL-1 positive cells began to appear in the matrix surrounding pulmonary endoderm. With the elongation of the lanryngo-tracheal groove in the direction of aortic sac from ED 10 to ED 11.5, ISL-1 positive prepharyngeal cells formed a cone-shaped structure centered by pulmonary endoderm, and its ventral end protruded into the cavity of aortic sac to form aorto-pulmonary septum. During the development of lanryngo-tracheal groove, a solid endoderm cord could always be observed at the ventral end of lanryngo-tracheal groove, and pulmonary endoderm of lanryngo-tracheal groove and its solid cord, to which strong Ptc1 and Ptc2 expression were mainly confined, was located in the center of the ISL-1 cone-shaped structure. From ED 12 to ED 13，separation of foregut at the level of outflow tract led to the formation of trachea, endoderm cord no longer be found and Ptc1 and Ptc2 positive expression disappeared in the tracheal epithelial. The density of the ISL-1 positive mesenchyme cells reduced sharply in the ventral to trachea, which gradually stopped to be added to outflow tract, and aortic sac was separated eventually.  Conclusion The differentiation and development of pulmonary endoderm are closely associated with the ISL-1 positive prepharyngeal mesenchyme of second heart field aggregation. The activity of sonic hedgehog(SHH) signaling system is in a high level during the pulmonary endoderm development, the developing pulmonary endoderm probably acts as an organizing center via SHH pathway to induce aggregation of ISL-1 positive cells, and the mechanical tension produced by endoderm growth is also to drive those ISL-1 positive cells migration for the normal morphogenesis of outflow tract. References | Related Articles | Metrics

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Preparation and histological evaluation of the decellularized scaffold for porcine small intestinal submucosa WANG Fu-yan DU Li-qun* 2016, 47 (6):  824-828.  doi: 10.16098/j.issn.0529-1356.2016.06.018 Abstract ( )   Objective To evaluate the different concentrations of sodium dodecyl sulfonate（SDS）on efficiency of cell removal from porcine small intestinal submucosa（SIS）and optimize the best concentration to produce an acellular porcine SIS scaffold for use in developing a tissueengineered corneal epithelium. Methods Fresh porcine SIS were decellularized with SDS (concentration of 0.1%, 0.2%, 0.3%, and 0.5% for 15 minutes, 30 minutes, 1 hour, and 2 hours) and randomly divided into 4 groups (A, B, C, D, 20 pieces in each group ) and the decellularization process was observed at the same time.After decellularization，the scaffolds were examined through the HE, 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) staining and the genomic DNA contect analysis to observe the biological characteristics of SIS. Results A cellular SIS was shown ivory, translucent and had certain transmittance. HE and DAPI staining revealed that when treatment with the time of 30 minutes in group A and B SIS were decellularized completely and preserved the overall tissue histoarchitecture, the collagen fiber porosity was increased, the acellular rate was more than 90%. Conclusion The acellular matrix treatment effect of 0.1% and 0.2% SDS for a period of 30min is satisfactory. The immunogenicity of the SIS can be reduced, therefore, it is an ideal acellular concentration for SIS and provides a theoretical basis for the construction of tissue engineering corneal epithelium. References | Related Articles | Metrics

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Role of Reelin signaling pathway in the pathogenesis of Alzheimer’s disease model mice YAN Ming-chao ZHAO Pei-wen CAO Jing-jing WANG Xiao-qing WANG Qian SUN Yi-zheng CHENG Yan-hong DENG Jin-bo* 2016, 47 (6):  721-730.  doi: 10.16098/j.issn.0529-1356.2016.06.001 Abstract ( )   Objective To investigate the role of DNA methylation, Reelin and Notch pathways in the pathogenesis of Alzheimer’s disease(AD). Methods A total of 184 wild-type (WT) and AD mice were used for Golgi staining, immunofluorescent labeling, Western blotting and transmission electron microscopy in this study. Results The pathological changes occurred after postnatal 6 months (P6M) in the hippocampus and neocortex of AD mice, such as amyloid plaques and neurofibrillary tangles. Astrocytes and microglia usually were clustered around amyloid plaques in AD mice. With age increasing, Reelin accumulated around amyloid plaques and formed amyloid-like deposits. In AD mice, the expression of Notch1 receptor with both full-length Notch1 and Notch intracellular domain(NICD) decreased. DNA methylation decreased in AD hippocampus and neocortex; especially in amyloid plaques, the DNA methylation almost disappeared. For instance, the expression of Dnmt3a and Dnmt1 were decreased in AD mice. Conclusion The amyloid plaques in AD mice can induce the cluster of astrocytes and microglia and the accumulation of Reelin. The abnormal expressions of Notch1 receptor and DNA methyltransferase probably are the causes of the neural dysfunction in AD. References | Related Articles | Metrics

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Anti-aging effects of angelica sinensis polysaccharides on brain aging induced by D-galactose in Nestin-green fluorescent protein transgenic mice and its mechanism YAO Hui CHEN Lin-bo CHEN Xiong-bin LIU Ying XIA Jie-yu WANG Zi-ling XIANG Yue XIONG Li-rong WANG Lu WANG Shun-he* WANG Ya-ping* 2016, 47 (6):  731-737.  doi: 10.16098/j.issn.0529-1356.2016.06.002 Abstract ( )   Objective To explore the anti-aging effects of angelica sinensis polysaccharides(ASP) on brain aging induced by D-galactose in Nestin-green fluorescent protein (GFP )transgenic mice and its mechanism. Methods Male Nestin-GFP transgenic mice (n=40) aging from 6 to 8 weeks old were randomly divided into the normal, ASP normal, brain aging model and ASP brain aging model groups. The brain aging model group was subcutaneously injected with D-galactose (200 mg/kg), qd×42 days. The ASP brain aging model group was intraperitoneally injected with ASP (140 mg/kg) since the 16th day of the replication in the brain aging model, qd×27 days. The ASP normal group was subcutaneously injected with the same amount of saline, qd×15 days, and following intraperitoneally injected with ASP (140 mg/kg) qd×27 days. The normal group was subcutaneously injected with an equal volume of saline within the same time. The related experiment was performed on the second day after finishing copying the model. Learning and memory abilities were measured by Morris water maze. Frozen sections were made to observe the hippocampus fluorescence intensity. The number of senescent cells were detected by senescence-associated β-galactosidase (SA-β-Gal) staining. The activities of superoxide dismutase (SOD), total antioxidant capacity (T-AOC) and contents of malonaldehyde (MDA) in hippocampus were quantified by chromatometry. The level of interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α proinflammatory cytokines in hippocampus were detected by methods enzyme-linked immunosorbent assay (ELISA). Results In the brain aging model group, the spatial learning and memory capacities were weaken, the fluorescence intensity decreased in the dentate gyrus (DG) of hippocampus, SA-β-Gal positive granules increased in section of brain tissue，and the activity of SOD, T-AOC decreased in hippocampus while the contents of MDA increased in hippocampus. The level of IL-1β, IL-6 and TNF-α increased in hippocampus. Compared with the brain aging model group, the spatial learning and memory capacities in the ASP brain aging group, were enhanced, the fluorescence intensity increased in the DG area of hippocampus, SA-β-Gal positive granules decreased in section of brain tissue, and the activity of SOD, T-AOC increased in hippocampus while the contents of MDA decreased in hippocampus. The level of IL-1β, IL-6 and TNF-α in hippocampus decreased. Conclusion ASP can antagonize brain aging induced by D-galactose in mice. In addition, improvement of antioxidant ability, down regulation of the level of proinflammatory cytokines and maintaining the number of neural stem cells in hippocampus may be the underlying anti-aging mechanism of ASP.  References | Related Articles | Metrics

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Change of learning and memory ability in neonatal rats with hypoxic-ischemic white matter injury LIN Ling ZHANG Geng LIN Qiao-mei LIN Qing* 2016, 47 (6):  738-743.  doi: 10.16098/j.issn.0529-1356.2016.06.003 Abstract ( )    Objective To explore changes of the learning and memory ability as well as the myelin basic protein (MBP) expression during various growth periods in neonatal rats with hypoxic-ischemic white matter injury (WMI). Methods New born 4-day-old SD rats were randomly divided into a control group and a hypoxicischemic WMI group. The histology of rat brain tissues was observed by HE staining at 4w after operation. The neurobehavior and the ability of learning and memory were investigated at 4weeks, 8weeks and 12weeks after operation. The distribution and expression of MBP protein were tested by immunofluorescent histochemical staining method and western blotting. Results The hypoxic-ischemic white matter injury was showed by HE staining in WMI group. The weaker ability of neurobehavior and learning and memory were displayed in WMI group compared with the control group (P＜0.01). The expression of MBP was significantly decreased in the WMI group compared with the control group（P＜0.01）. Conclusion The ability of learning and memory was weakened and the expression of MBP protein was decreased in new born 4-day-old SD rats with hypoxic-ischemic white matter injury. The weaker ability of learning and memory may be resulted from the decreased expression of MBP protein. References | Related Articles | Metrics

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Effect of Tenuigenin on low density lipoprotein receptor-related protein 1 level changes of Alzheimer’s disease rat in hippocampus NI Jie FANG Yu HU Yan DONG Yu-lin* 2016, 47 (6):  744-749.  doi: 10.16098/j.issn.0529-1356.2016.06.004 Abstract ( )   Objective To observe the effect of Tenuigenin (TEN) on low density lipoprotein receptor-related protein 1（LRP1）level in Alzheimer’s disease (AD) rat hippocampus. Methods Sixty SD rats were randomly divided into groups respectively. β-Amyloid protein (Aβ)1-40 was injected into hippocampus CA1 region to establish AD model by brain stereotactic technique and then rats were treated with intragastric TEN for four weeks. Learning and memory abilities were assessed by Morris water maze and the Real-time PCR and Western blotting were used to analysis the expression of LRP1 mRNA and protein. The changes of LRP1 positive neurons were checked by immunofluorescence staining. Results 1. Morris water maze: the learning and memory ability of the AD group was significantly decreased than that of the normal group（P<0.01）.After TEN gastric lavage treatment, the escape latency was shortened, the swimming distance was reduced, and the frequency of the virtual platform was increased in every TEN treatment group, there were a statistically significant difference in middle and high dose group（P<0.05 or P<0.01）. 2. Real-time PCR and Western blotting: the relative expression levels of LRP1 mRNA and protein in AD group were significantly lower than that in normal control group (P<0.01). After TEN treatments, the relative expression of LRP1 mRNA and protein in the treatment group increased gradually with the increase of the dosage of TEN. The effect of low dose group was not significant (P>0.05), but there were a significant difference between middle and high dose groups (P<0.01). 3. Compared with normal group, the number of LRP1 positive cells in the AD group decreased significantly (P<0.01), but in the high dose group increased significantly (P>0.05). Conclusion TEN may improve the learning and memory ability of Aβ 1-40induced AD rats by increasing the content of LRP1 in the brain. References | Related Articles | Metrics

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Soluble overexpression and immunological activity of the Newcastle disease virus hemagglutinin-neuraminidase protein ZHANG Wei-qiang GUO Yu-jie LI Sai-sai WANG Yue-ying CHEN Li-ying YANG Guo-yu* 2016, 47 (6):  848-855.  doi: 10.16098/j.issn.0529-1356.2016.06.023 Abstract ( )   Objective To improve the soluble overexpression of Newcastle disease virus(NDV) hemagylutinin-neuraminidase(HN) protein in E. Coli and determine its immunological. Methods Ten plasmids that expressed NDV HN protein with-N-terminal Grifin, gultathione-S-transferase(GST), maltose biniding protein(MBP), NusA, sanall ubiquitin-related modifier(SUMO), thioredoxin, protein G, γ-crystallin, Ars C or Ppi B tags were constructed to test the effects of the various tags on the expression level and solubility of NDV HN protein in E. coli. The sequences of the ten plasmids were confirmed and the ten plasmids were transformed into E. coli BL21 strain. In order to select and screen the best expression conditions, different induced dose isopropy-β-D-thiogalactoside(IPTG) final concentration, culture temperature, Amp final concentration and culture media were used. The expression level and solubility of fusion protein were analyzed by SDS-PAGE and Western blotting. The fusion proteins were purified by Ni-NTA chromatography to develop indirect ELISA. The purified fusion protein was detected by Western blotting and indirect ELISA. Results Ten HN fusion protein expression plasmids were constructed successfully. Maltose binding protein(MBP) tag significantly increased the HN protein soluble overexpression. The expression level was the highest. The results from Western blotting and indirect ELISA showed that the purified HN protein had a good immunogenicity. Conclusion MBP tag enhances the expression and solubility of HN protein. The study will lay foundation for the research on the NDV mechanism, vaccine and diagnostic reagents. References | Related Articles | Metrics

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Medical image segmentation and creation of 3D models for embedding into 3D PDF files YANG Xiao-kai* WU Shu-zhi CHEN Xiao-su 2016, 47 (6):  856-858.  doi: 10.16098/j.issn.0529-1356.2016.06.024 Abstract ( )   Objective To discuss how to convert medical images into a 3D model and embed them in a Portable Document Format (PDF) file. Methods Three opensource programs, 3D Slicer,MeshLab,TeXstudio, were used to volume clipping with ROI for segmentation models and embed them in 3D PDF. Results 3D Slicer used a variety of methods for model segmentation,Meshlab carried out model cleaning and segmentationand, generated TEX format files, exported U3D format files,and then generated 3D PDF files that were compiled by TeXstudio. Conclusion The method introduced in this paper is simple and easy to learn, and is helpful to promote the production and use of 3D PDF. References | Related Articles | Metrics

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Effects of senescent bone marrow stromal cells on aging of hematopoietic cells CHEN Xiong-bin CHEN Lin-bo LIU Ying JING Peng-wei HOU Ji-ying XIA Jie-yu SONG Xiao-ying XIONG Li-rong WANG Lu WANG Ya-ping* 2016, 47 (6):  750-755.  doi: 10.16098/j.issn.0529-1356.2016.06.005 Abstract ( )   Objective To investigate the effect of senescent bone marrow stromal cells (BMSCs) on aging of hematopoietic cells. Methods The BMSCs were isolated by whole bone marrow adherent culture from the rat, and passaged to 3rd generation (P3). The P3 generation of BMSCs was collected and divided into two groups. The control group was cultured as routine and the aging group was cultured with additional 30g/L D-Galactose (D-Gal) for 48 hours. And then, the amounts of granulocyte macrophage colony-stimulating factor(GM-CSF), stem cell factor(SCF), interfeukin(IL)-6, IL-1β，IL-2 and tumor necrosis factor-α(TNF-α) in BMSCs culture supernatant were assayed by ELISA; the proliferation ability of BMSCs was detected by cell counting Kit-8(CCK-8); the senescenceassociated β-galactosidase（SA-β-Gal）staining was used to detect the senescent BMSCs. Bone marrow mononuclear cells(BMMNCs) from rat femur marrow were isolated and purified. Co-culturing aging BMSCs with BMMNCs for 24 hours, and then BMMNCs was collected. The proliferation ability of BMMNCs were measured by CCK-8; the proliferation and differentiation ability of CFU-Mix was detected by colony forming assay; the SA-β-Gal staining was used to detect the senescent BMMNCs; the cell cycle distribution and the ratio of apoptosis of BMMNCs were analyzed by flow cytometry (FCM); DCFH-DA fluorescent staining and FCM were analyzed the level of reactive oxygen species (ROS) in BMMNCs. Malondialdehyde (MDA) content and total superoxide dismutase (SOD) activity were analyzed using enzymatic assay. Results The proliferation of aging group BMSCs was significantly decreased. The positive ratio of SA-β-Gal was markedly increased. The amount of GM-CSF, SCF, IL-6, IL-1β in BMSCs culture supernatant of aging group were obviously decreased and the level of IL-2、TNF-α were raised. After co-cultured BMMNCs with aged BMSCs for 24 hours, the positive ratio of SA-β-Gal stained BMMNCs was significantly increased; the proliferation ability of BMMNCs were declined; the number of CFU-Mix formation was significantly decreased; the BMMNCs were held in G1 phase, and the apoptosis rate was increased; the level of ROS and MDA in BMMNCs was significantly increased and total SOD activity was inhibited. Conclusion D-Galactose may establish BMSCs aging in vitro. The senescence of BMSCs may lead to the hematopoietic cells dysfunction, and the mechanism may be related to the effect of oxidative damage. References | Related Articles | Metrics

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Inhibition of bone marrow mesenchymal stem cells on epithelial mesenchymal transformation induced by transforming growth factor-β1 WANG Xin GAO Jun-ling* ZHAO Man-man ZHU Hui-xing TIAN Yan-xia LI Ran JIANG Xiao-hua YU Lei TIAN Jing-rui CUI Jian-zhong 2016, 47 (6):  756-762.  doi: 10.16098/j.issn.0529-1356.2016.06.006 Abstract ( )   Objective To explore whether bone marrow mesenchymal stem cells could inhibit epithelial-mesenchymal transition(EMT) of rat type Ⅱalveolar epithelial cells through inhibiting Smad3 phosphorylation, and play a role on anti pulmonary fibrosis. Methods Rat type Ⅱ alveolar epithelial cell line RLE-6TN cells were cultivated, induced by 5μg/L transforming growth factor(TGF)-β1 in vitro, and divided into normal control group. TGF-β1 induced group and BMSCs intervention group. Morphological changes of epithelial-mesenchymal transition in RLE-6TN cells were observed under an inverted phase contrast microscope. Immunofluorescence methods were used to determine the location of E-cadherin (E-cad) and α-smooth muscle actin（α-SMA). The expressions of E-cad, α-SMA, p-Smad3 and Snail1 were detected by Western blotting in each group. Results The process of EMT induced by TGF-β1 was accompanied by morphological alteration. RLE-6TN cells had an epithelial cobblestone-like morphology, a round or polygonal shape and exhibited very close cell-cell proximity, while after exposure to TGF-β1, the cells displayed spindle-shaped elongated fibroblast-like cells morphology. Concomitant with a down-regulation of the epithelial markers E-cad, the expression of interstitial cells markers α-SMA up-regulated. Similar morphological changes were noticeably suppressed in the BMSCs intervention group, and the BMSCs intervention group showed a reduction in TGF-β1-induced up-regulation of α-SMA, while a down-regulation of E-cad was observed. Compared with control group, the expression levels of p-Smad3 and Snail1 increased significantly after the stimulation of TGF-β1. After BMSCs intervention,the expressions of p-Smad3 and Snail1 significantly decrease compared to the TGF-β1 group. Conclusion BMSCs may block TGF-β1/Smad3 signal transduction pathway, and then inhibit epithelial mesenchymal transition of type Ⅱ alveolar epithelial cells. References | Related Articles | Metrics

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Effect of the Hedgehog signaling pathway blockers cyclopamine on the articular cartilage cell proliferation of adjuvant arthritis rat in vitro SONG Xian-bing ZHANG Jun-qiang CAO Wei YANG Xiao-si LIU Mei-mei CHEN Xiao-yu* 2016, 47 (6):  763-768.  doi: 10.16098/j.issn.0529-1356.2016.06.007 Abstract ( )   Objective To explore the effect of Hedgehog (Hh) signaling pathway inhibitor cyclopamine on articular cartilage cell proliferation and part of the mechanism in adjuvant arthritis (AA) rats modelin vitro. Methods Freund’s complete adjuvant induced AA rats, measurement of arthritis index and secondary foot swelling degree, rat’s cartilage tissue growth were observed by HE staining;AA rat ankle joint cartilage, pancreatic enzyme-collagen enzymatic isolation, culture, identification, AA rat ankle joint cartilage cell proliferation was determined by MTT method. Cyclopamine (0, 0.05, 0.5, 5, 20 μmol/L) in vitro, cell apoptosis was determined by Annexin V-FITC/PI double dye in AA rats ankle joint cartilage, proteins expressionin in AA rats ankle joint chondrocytes of Shh, Ptch1, Gli1 were determined by Western blotting. Results After freund's complete adjuvant induced, compared with normal rats, the AA rats arthritis index and secondary foot swelling degree increased significantly, HE staining showed AA rats ankle cartilage destruction; Toluidine blue and type Ⅱ collagen in vitrocultivation of AA rats ankle joint cartilage cells; Cyclopamine (0.05, 0.5, 5, 20 μmol/L) in vitro could elevated AA model of articular cartilage cell proliferation. Flow cytometry test results showed that cyclopamine reduced apoptosis rate of AA model of chondrocytes; Compared with the model group, cyclopamine (0.5, 5, 20 μmol/L) declined AA chondrocytes related proteins express (Shh, Ptch1 and Gli1) of Hh signaling pathway significantly. Conclusion AA model rat was established by induced Freund’s complete adjuvant. Cyclopamine may inhibit the proliferation of chondrocytes in AA rats in vitro, inhibit the apoptosis of cartilage cells, which was associated with inhibition of AA rat chondrocytes Hh signal. References | Related Articles | Metrics

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Analysis of bone sialoprotein and mature peptide modification sites and evolutionary trace CAI Min LIN Jian-li* HOU Jian-ming TANG Fa-qiang JIN Long 2016, 47 (6):  769-773.  doi: 10.16098/j.issn.0529-1356.2016.06.008 Abstract ( )   Objective To investigate the expression of osteoblasts bone sialoprotein(BSP) and evolutionary trace. The mature peptide gene sequences were analyzed to obtain BSP modification sites. Methods Total RNA was isolated from cultured osteobalsts and desired cDNA fragment was obtained by RT-PCR with two gene specific primers. The segment was inserted into pBluescript KS vector and the result plasmid was transformed into DH5α. The positive clone and sequence were performed. Using BSP as seed sequences, the searching of BSP gene sequence and its homologous protein with various bioinformatics tools were retrieved from NCBI database. The comparative studies were taken for BSP evolutionary trace sites and related functional sites. Results There was a complete structural domain of 72 homologous protein sequences, 16 bone sialoprotein family BSP-propeptide domain, 10 RGD binding domain and the BSP domain 33 subfamily specific residues. The ligand of bone sialoprotein RGD domain binding sites was mainly distributed in the middle region with pockets. 900bp specific fragment was obtained. The sequence of the gene encoding BSP mature pepide coincided with that of the references published. Conclusion The gene encoding BSP mature peptide is obtained from osteoblasts that express and translate into active bone sialoprotein. There are methylation and acetylation sites existed on the BSP mature peptide. This result will help us to further investigate the expression and function of BSP. References | Related Articles | Metrics

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Correlation between stature and hand length, hand breadth, foot length and foot breadth in Muya WANG Wen-jia ZHANG Xing-hua* YU Ke-li YAN Shi BAO Jin-ping YANG Ya-jun WANG Zi-shan ZHENG Lian-bin 2016, 47 (6):  829-832.  doi: 10.16098/j.issn.0529-1356.2016.06.019 Abstract ( )   Objective To investigate the relationship between stature and hand length, hand breadth, foot length and foot breadth of Muya in Sichuan Province, in order to provide a scientific basis for anthropology, forensic medicine and clinical medicine and to accumulate data for the Chinese constitution investigation. Methods A methodology and a standard of Anthropometry recognized were used to measure stature and hand length, hand breadth, foot length and the foot breadth of 157 aged 20-70 adults (77 males and 80 females ) of Muya in Sichuan province. Results The mean values of stature,hand length, hand breadth, foot length and foot breadth, as well as the correlation coefficient were caculated. Ther value of each correlation coefficient was between 0.279-0.623. The relationships of the hand length and stature, hand breadth and stature, foot length and stature, foot breadth and stature were correlated, ther values were: male 0.285, female 0.345,male 0.500, female 0.379, male 0.623, female 0.620, male 0.359 and female 0.279 respectively. Conclusion The eight caculated regression equations are eligible for evaluating the stature of Muya in Sichuan Province. References | Related Articles | Metrics

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Difference of 25 biological phenotypes between the left and right sides of middle aged residents in Nanyang BAI Yong-gang WANG Yan-ke FAN Ying XU Guo-chang* LIU Rong-zhi 2016, 47 (6):  833-837.  doi: 10.16098/j.issn.0529-1356.2016.06.020 Abstract ( )   Objective To investigate the biological characteristics of middle aged people in Nanyang city, and to compare the left and right differences between them. Methods Twenty-five biological phenotypes of 495 healthy urban residents aged 45-59 years (230 males and 265 females) were collected and analyzed statistically using the method of physical fitness measurement and the intuitive determination. Results The right of the upper limb length, arm length, forearm circumference, grip, pedal strength, a-b ridge count in the men and women were higher than the left（P＜0.05）, while the length of foot, leg circumference, triceps skinfold thickness were less than the left（P＜0.05）. Finger ridge count (FRC), percent distance of axial triradius (tPD) in the male index of the right side were higher than the left. Systolic blood pressure, thigh skin fold thickness, and tPD left of the female were greater than right side（P＜0.05). Arm circumference, and FRC of the little finger of the female were greater than the left（P＜0.05）. Other indicators were not statistically different. Conclusion There is a left and right side difference in the biological characteristics of the upper and low limbs of the middle aged people in Nanyang. The data provide a baseline information for the whole picture of the physical characteristics of the population. References | Related Articles | Metrics

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Characteristics of bone strength index and its relationship with body composition in Lanzhou Han nationality adults QIN Qing-song WANG Dong-mei MA Lu-tong YU Hua SUN Xue-mei MA Li-rong BAI Jing-ya* 2016, 47 (6):  838-842.  doi: 10.16098/j.issn.0529-1356.2016.06.021 Abstract ( )   Objective To analyze the change of bone strength index (SI) and its relationship with body composition (BC) in Lanzhou Han nationality adults and to provide reference data for prevention and diagnosis of osteoporosis (OP). Methods Ultrasonic bone mineral density meter and Bioelectric impedance technique were used to test the bone strength index of right calcaneus and body composition. A total of 2291 Han nationality adults in Lanzhou（1021 males,1270 females) were sampled. The association between bone strength index and body composition was tested by multiple linear regressing analysis. Results The bone strength index of Lanzhou Han nationality adults presented tendency of decrease along with age, and the peak of bone mass (PMB) occurred at 20-30 years old. The SI in male decreased rapidly between 20-30 and 50-60 years old, and the SI in female fell fast beyond 50 years old. The SI in male was higher than female at each age group. Compared with other nationalities by using the same method, the mean values of SI and PMB were lower in Lanzhou Han nationality adults and PMB in Lanzhou Han female, respectively. According to the analysis of multiple linear regressing between SI and BC，both muscle mass and subcutaneous fat mass, and body mass index(BMI) were positively associated with SI, visceral fat mass was negatively correlated with SI. Conclusion The SI and PMB values are respectively lower in Lanzhou Han nationality adults and Lanzhou Han female. The PMB appeares early in youth. Females above 50 years old are a higher risk population of OP. Muscle mass, subcutaneous fat mass and BMI are protective factors for OP while visceral fat mass is a factor of risk for OP. References | Related Articles | Metrics

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Verification of Allen’s rule: physical data from Chinese rural Han LI Yong-lan* 2016, 47 (6):  843-847.  doi: 10.16098/j.issn.0529-1356.2016.06.022 Abstract ( )   Objective To focus on the variation rule of morphological characteristics of rural Han Chinese adults in districts of different mean annual temperature, and to try to verify whether they were consistent with Allen’s Rule. Methods Sixteen indicators including statures and body weights of 16,501 cases of Chinese rural Han adults in 36 districts across China were measured from 2009 to 2013 and 6 indices were calculated. Results Upper extremity length, forearm length and SUELI were positively correlated with the mean annual temperature. Maximum forearm circumference and FLGI (and UALGI of female) were negatively correlated with the mean annual temperature. The larger of thigh of male made their lower limbs larger as mean annual temperature went down. Their lower limbs were a little more stout. The length and circumferences of lower limb of female had no significant change when mean annual temperature went down. Conclusion As mean annual temperature goes down, the change of upper limb of Chinese rural Han is consistent with Allen’s Rule, and the change of lower limb is not consistent with Allen’s Rule. References | Related Articles | Metrics

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Role of hypoxia-inducible factor-1 in immune escape of tumor cells XIONG Wei-peng* 2016, 47 (6):  859-961.  doi: 10.16098/j.issn.0529-1356.2016.06.025 Abstract ( )   Hypoxia is a hallmark of tumor, and hypoxia-inducible factor (HIF-1) is an essential regulator of hypoxia adaptation in tumor progression. Hypoxia and HIF-1 signaling are important components of the tumor microenvironment that modulates the tumor immune response. Tumor hypoxia via HIF-1 signaling enables tumor cell to develop mechanisms to evade from immune surveillance and attack. This mini-review emphasizes on the molecular mechanisms of HIF-1-mediated tumor cell escape and the development of inhibitors targeting HIF-1. References | Related Articles | Metrics

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Susceptibility-weighted imaging of deep medullary veins CHEN Zheng-zhen QIAO Hui-huang GUO Yu REN Chuan-gen ZHANG Xiao-fen LI Jian-ce CHEN Cheng-chun* 2016, 47 (6):  796-801.  doi: 10.16098/j.issn.0529-1356.2016.06.013 Abstract ( )   Objective To investigate the caliber, length, distribution and drainage of deep medullary veins by using susceptibility-weighted imaging (SWI). Methods Sixty healthy volunteers were examined using SWI on a 3.0T magnetic resonance system. The raw data were transferred to the Extended MR Workspace 2.6.3.4 workstation. The SWI images were reconstructed by using minimum intensity projections (mIPs) technique in the transverse plane. The SW images were combined with the corresponding images of T1WI by using Photoshop CC (Adobe Systems, USA) to visualize the positional relationship between the course of veins and the cerebral structures. Results Firstly, the caliber of deep medullary veins was basically same and ranged from 0.2-0.3mm. Secondly, according to the position, the deep medullary veins were divided into three area: the anterior, central and posterior. Thirdly, the incidence of the deep medullary veins ranged from 4-10 in the anterior area, 8-19 in the central area, 3-7 in the posterior area. Fourthly, the deep medullary veins in the central area were longer than other areas. Finally, the deep medullary veins in the anterior, central, posterior area were respectively drained into the anterior septal vein and anterior caudate vein, the transverse caudate vein, and the medial atrial vein. Conclusion Our study suggests that SWI of the deep medullary veins is feasible to create the cerebral medullary venous network without using a contrast agent and provides more theories to the classification and mechanism about abnormality of deep medullary veins. References | Related Articles | Metrics