Loading...

Table of Content

    2015, Volume 46 Issue 4
    06 August 2015
    Previous Issue    Next Issue

    For Selected: Toggle Thumbnails
    Body adiposity index values of Chinese urban Han adults distributed in different latitude areas
    LI Yong-lan ZHENG Lian-bin* XI Huan-jiu YU Ke-li
    2015, 46 (4):  572-576.  doi: 10.16098/j.issn.0529-1356.2015.04.022
    Abstract ( )  

    Objective To study the changes of body adiposity index (BAI) of Chinese urban adult with latitude. Methods Based on the measurement of stature and hip circumference of 10 451 Chinese urban Han of 31 districts form 2009 to 2013, and calculated their body adiposity index values, BAI. Results The average of BAI of 31 male groups was between 23.64 and 27.49. It was divided into 3 levels:the first level was BAI≥26.00, the second level was between 25.00 and 25.99, and the third level was BAI<25.00; The average of BAI of 31 female groups was between 27.56 and 30.80. It was divided into 3 levels: the first level was BAI≥30.00, the second level was between 29.00 and 29.99, and the third level was BAI<29.00. Conclusion Hip circumference has a positively linear correlation with latitude,and stature has a positively linear correlation with latitude, hip circumference and stature have positively linear correlation, BAI has positively linear correlation with hip circumference and latitude, but BAI and stature hare no correlation of 31 urban Han groups.

    References | Related Articles | Metrics
    Expression of dysbindin-1 during spermatogenesis
    YAN Hui LIU Hui-xia WEI Zong-bo GUO Rui ZHANG Tao FENG Ya-qin*
    2015, 46 (4):  548-552.  doi: 10.16098/j.issn.0529-1356.2015.04.018
    Abstract ( )  

    Objective To test dysbindin-1 expression during spermatogenesis and dysbindin-1 effect on histology of acrosome in mice. Methods Western blotting analysis was applied to detect the expression of dysbindin-1 in testis on day 7,14,21,28 and 35 postpartum and three-month old mice(n=3). The caudal spermatozoa was collected from sdy mice (no dysbindin-1expression), HE staining was performed for sperm morphology, fluorescein isothiocyanate conjugate-pisum sativum agglutinin (FITC-PSA) staining and immunofluorescence staining with antibody against sperm protein 56(sp56) for acrosome on sperm smear. Results Dysbindin-1A, not -1C, was expressed in postnatal testis development and spermatozoa. Mature sperm from sdy caudal epididymis had normal formation. Conclusion Dysbindin-1A, expressed in testis tissue and sperm during spermatogenesis, is not required for maintaining the morphology of sperm.

    References | Related Articles | Metrics
    Morphological study of Reelin regulation on the development of rostral migratory stream in the mouse
    SHI Shu-qin CUI Zhan-jun YAN Ming-chao MU Xiao-yun DENG Jin-bo* YIN Ming-wei*
    2015, 46 (4):  433-442.  doi: 10.16098/j.issn.0529-1356.2015.04.001
    Abstract ( )  

    Objective To investigate the differentiation of neural stem cells in the subventridular zone (SVZ) and neural migration along the the rostral migratory stream (RMS) into the olfactory bulb(OB), particularly, Reelin’s effect on the neural differentiation and migration in RMS. Methods Wild type mice (50 cases) and reeler mice (23 cases) from E16 (embryonic day 16) to P90 (postnatal day 90) were used for Nissl staining, immunofluorescent labeling, ink perfusion and ultrastructure observation to visualize neural stem cells, the radial glial cells and vasulature in neocortex and RMS. The development of RMS in two groups was compared. Results In embryonic days and early postnatal period, vascular niches in the SVZ consisted of neurons, glial cells, neural stem cells and vascular vessel which provided a ideal microenvironment for the differentiation and mturation of neural stem cells. After maturation in the vascular niche, the neural stem cells started to migrate tangentially along RMS toward olfactory bulb with the chain-guidable pattern along both glial and vascular scaffold. After entering in OB, the migrating pattern translated into the radial migration, and these cells differentiated into various neuron in olfactory bulb. In general, RMS’s development, neural stem cell differentiation and cell migration in reeler mice were similar to the wild type, howover, the distribution of vasculature in cortex and RMS was disordered, and the migrating cells often concentrated in olfactory bulb densely and scatteredly. Conclusion Niche in subventricular zone is the main sources of neural stem cells, and both vasculature and radial glia and acts as a scaffold for the migration of neural stem cells in RMS. As an important signal for cell migration, Reelin can affect the development of blood vessels and cell migration in RMS. Without Reelin, the cell migration in RMS will be blocked.

    References | Related Articles | Metrics
    Morphological changes and expression of nerve growth factor of hippocampus in different ages of Bactrian camels
    ZHANG Yuan XIE Li WANG Chang-jiang YUE Ming-xia WANG De-gui SHAO Bao-ping WANG Jian-lin*
    2015, 46 (4):  443-449.  doi: 10.16098/j.issn.0529-1356.2015.04.002
    Abstract ( )  

    Objective To investigate morphological changes of neurons and the expression of nerve growth factor (NGF) in different areas of hippocampus in fetal, young and adult Bactrian camels. Methods Nissl staining was used to observe morphological changes of pyramidal cells in CA1-CA3 areas and granule cells of dentate gyrus. Immunohistochemical method was used to detect the expression of NGF of hippocampus in fetal, young and adult Bactrian camels. Results The results of Nissl staining showed that from fetal Bactrian camel to young Bactrian camel and to adult Bactrian camel, pyramidal cells in CA1-CA3 areas and granular cells in teh dentate gyrus enlarged in volume but decreased gradually in density with the growth of Bactrian camel. The proportion of nuclei in neurons also decreased. Immunohistochemical analysis revealed that the expression level of NGF in pyramidal cells of adult camel was significantly higher than the other two periods of Bactrian camel (P<0.01). Conversely, the expression level of NGF in granule cells increased consistently from fetal camel to adult camel. Conclusion From fetal Bactrian camel to adult Bactrian camel, the main neurons of hippocampus continuously become mature: neurons change from small and global cells which have little cytoplasm, to neurons which are large and have specific morphology and function. The high expression level of NGF in hippocampus of adult camel indicates that NGF could regulate differentiated neurons, propel morphological and functional changes in neurons, which will consequently contribute to the improvement of learning and memory functions of hippocampus in Bactrian camel.

    References | Related Articles | Metrics
    Morphological characteristics of atrioventricular endocardial cushion development of the mouse embryonic heart
    YANG Yan-ping JING Ya* HONG Xiao-yang LI Hai-rong QIAO Cong-jin WU Shan-shan
    2015, 46 (4):  543-547.  doi: 10.16098/j.issn.0529-1356.2015.04.017
    Abstract ( )  

    Objective To investigate the morphological characteristics of the formation and the fusion pattern of atrioventricular endocardial cushion of mouse embryonic heart.
    Methods Serial sections of thirty mouse embryos during embryonic day(ED) 9 to ED13 were stained immunohistochemically with antibodies against α-smooth muscle actin (α-SMA), myosin heavy chain (MHC) and phosphorylated histone H3 (PHH3). Semithin sections and ultrathin sections of fifteen mouse embryonic hearts from ED12 to ED13 were observed by light microscope and transmission electron microscopy. Results At ED9, the atrioventricular canal was observed between the atrium and the ventricle. The cardiac jelly layer was thickened and contained a few mesenchymal cells. At ED10, the atrioventricular endocardial cushion began to form. The inferior endocardial cushion was thicker than the superior cushion. The α-SMA and MHC positive myocardial cells neighboring with the inferior cushions extended towards the cushion. The mesenchymal cells in the cushions showed α-SMA or PHH3 negative. During ED11 to ED12, the inferior and superior cushions became symmetric and the mesenchymal cells increased obviously. Only a few of mesenchymal cells expressed α-SMA or PHH3. At ED12 and ED13, the bilateral cushions closed to each other and began to fuse. The cellular membrane touched each other and formed intercellular junctions between some mesenchymal cells. The mesenchymal cells contained rough endoplasmic reticulum and mitochondria, and few of microfilament were observed in the cells. Conclusion The first change of atrioventricular endocardial cushion formation of mouse embryonic heart is the morphologic change of the endothelial cell from squamous to cuboidal. The formation of the two endocardial cushions is not simultaneous. During the cushion fusion, cellular junction is formed locally between mesenchymal cells which contain few of microfilament. Thus ultrastructure characteristics of the fusion of the atrioventricular endocardial cushion is different with the outflow tract ridge.

    References | Related Articles | Metrics
    Generation of induced pluripotent stem cells derived from parthenogenetic embryonic stem cells and reprogramming of imprinted genes
    SHAN Zhi-yan WU Bin ZHANG Yue XUE Yuan WU Yan-shuang SHEN Xing-hui LEI Lei LIU Zhong-hua*
    2015, 46 (4):  553-557.  doi: 10.16098/j.issn.0529-1356.2015.04.019
    Abstract ( )  

    Objective To investigate the effect of pluripotency and the expression of imprinted genes on the parthenogenetic embryonic stem cells (PgESCs) by induced pluripotent stem cells (iPSCs) reprogrmming. Methods In our study, we firstly generated the PgESCs from parthenogenetic blastocyst, induced the pluripotent factors into the PgESCs by retrovirus transfection and generated the iPS cells derived from PgESCs. Results It showed no difference in pluripotency and differentiation between PgESCs and PgESC-iPS cells, however, the expression of maternal imprinted genes was significantly increased and the expression of paternal imprinted genes was decreased in PgESC-iPS cells compared to that in PgESCs. Conclusion These results demonstrated the expression of imprinted genes could be modified by iPSCs reprogramming, which may be close to the expression level of imprinted genes in fertilized embronic stem cells.

    References | Related Articles | Metrics
    Morphologcal study of the pecten oculi from ostrich (Struthio camelus)
    WANG Kai YANG Yu-rong FENG Yong-jie JIAO Xi-lan LIANG Hong-de*
    2015, 46 (4):  450-454.  doi: 10.16098/j.issn.0529-1356.2015.04.003
    Abstract ( )  

    Objective To investigate the anatomy and histology of 10 ostrichs’ pecten in order to provide morphological base for research of ostrich pecten oculi. Methods The morphology of the pecten oculi was observed with a microscope in the 10-month-old healthy Africa black-neck ostrich (Struthio camelus) by hematoxylin-eosin staining, Gordon-Sweet staining, and Van Gieson staining. Results Gross-anatomically, the pecten oculi was attached to the optic disc, and had an irregular cone shape and projected into the vitreous body. In the steromicroscope investigation, the pecten oculi was composed of 1 primary lamella, 18-22 secondary lamellae and a few tertiary lamellae. The primary lamella was vertically located in the central part of the pecten, the secondary lamellae were peripherally originated from the primary lamella with the tertiary lamellae which arose from the distal part of some secondary lamella. The pecten consisted of loose connective tissue and was rich in collagen fibers, blood vessel and pigment. Conclusion Pecten oculi in the ostrich is an ideal model for the research of the function of retinal blood vessel form mammal and human.

    References | Related Articles | Metrics
    Effects of zinc on P53 protein expression in hippocampal neurons of mice with spinal nerve transection -induced neuropathic pain
    WANG Yue-jing ZHANG Hai-yan LIAN Jie YAO Hong-bo SUN Li-hui* ZHANG Meng
    2015, 46 (4):  455-461.  doi: 10.16098/j.issn.0529-1356.2015.04.004
    Abstract ( )  

    Objective To investigate the effects of the zinc on learning,memory and the expression of P53 in the hippocampus of mice with L5 spinal nerve transection -induced neuropathic pain(NPP). Methods Forty-eight adult male mice were randomly divided into 3 groups (n=16 per group): control group, NPP group and high zinc NPP group (zinc-NPP group). In the control group, the L5 spinal nerve was exposed but without ligation. In the NPP group, the nerve was ligated on the left. The mice of the zinc - NPP group drank a zinc sulfate solution after the left L5 spinal nerve ligation. After 29 days, the spatial learning and memory ability in three groups mice were measured by Morris water maze experiment. After the experiment was finished, cell apoptosis rate was detected by using flow cytometry,P53 expression of each group was tested by immunohistochemical and immune imprinting technology. Results The incubation period of finding the hidden platform was gradually shortened in the NPP group(21.03±12.08),zinc-NPP group(16.97±8.59)and control group(8.74±3.61) (P<0.05). The proportion of the time and the path to the total time and the total path of finding the Hidden platform gradually increased in NPP group, Zn-NPP, and control group (P<0.05). Flow cytometry detection results shown that the apoptosis rate of Control group(1.88%),Zn-NPP group(16.12%)and NPP group(33.17%)gradually increase(P<0.05).The expression of P53 in the hippocampus CA1 area of the control group,Zn-NPP group and NPP group was 27.42±7.08, 37.52±10.58, and 53.31±9.26, respectively.The expression of P53 in hippocampus CA3 area of the control group,Zn-NPP group and NPP group was 29.51±8.31, 36.21±7.91, and 56.71±8.09, respectively. The expression of P53 in the hippocampus of the control group,Zn-NPP group and NPP group by immune imprinting technology was 29.51±8.31, 36.21±7.91, 56.71±8.09, respectively. All the result showed a gradually increasing trend. Conclusion Zinc can improve the ability of learning and memory in neuropathy pain mice.

    References | Related Articles | Metrics
    Expression of vitamin A in peripheral blood and differential expression of retinoic acid receptor α and HA117 in colon segments of children with Hirschsprung’s disease
    LUO Yuan-yuan TENG Yin-ping LIU Hang LI Li JIN Xian-qing*
    2015, 46 (4):  558-563.  doi: 10.16098/j.issn.0529-1356.2015.04.020
    Abstract ( )  

    Objective To describe the expression profiles of vitamin A in blood serum and retinoic acid receptor α(RARα), HA117 in different portions of the colon of patients diagnosed with Hirschsprung’s disease (HD). Methods Peripheral blood and colon specimens were collected from 17 HD patients. The concentration of vitamin A was detected in blood serum, and the colon specimens were grouped into 3 segments: anastomosis, dilated segment and stenotic segment. Levels of RARα mRNA and HA117 RNA were evaluated by Realtime PCR. RARα protein expression were analyzed by immunohistochemistry and Western blotting. Results The concentration of vitamin A in peripheral blood is (0.73±0.05) μmol/L, which was below the normal level. The levels of RARα mRNA expression was significantly lower in the stenotic segment compared to proximal anastomosis (P<0.05,n=17). The level of HA117 RNA expression was significantly higher in the stenotic segment compared to proximal anastomosis (P<0.001,n=17). RARα were expressed in ganglion cells and nerve fibers between the circular and longitudinal muscles in anastomosis; positive staining was diminished in the stenotic segments. The levels of RARα protein expression was significantly lower in the dilated segment(P<0.05,n=17)and stenotic segment(P<0.001, n=17)compared to proximal anastomosis. Conclusion The vitamin A and RARα may contribute to the development of enteric nervous system (ENS); long non-coding RNA (LncRNA) HA117 may have an anti-differentiation function, and play a pivotal role in the progression of HD.

    References | Related Articles | Metrics
    Extramuscular nerve block targets localization for the treatment of spasticity of the medial group of the muscles of the thigh
    HU Shuang-jiang TIAN Xu-feng ZHANG Yong YANG Sheng-bo *
    2015, 46 (4):  532-537.  doi: 10.16098/j.issn.0529-1356.2015.04.015
    Abstract ( )  

    Objective To determine the precise location of extramuscular nerve block targets of thigh adductor muscles. Methods A horizontal reference line (line H) was designed from the pubic tubercle to the skin lateral to the greater trochanter of the femur. Another longitudinal reference line (line L) was designed from the pubic tubercle to the level of the medial epicondyle of the femur. The obturator nerve and its muscular branches were dissected and exposed in 10 formalin fixed adult cadavers (20 sides). The nerves were coated barium sulfate on the surface and air-dried, X-ray radiography was performed. The percent positions of extramuscular nerve block targets of muscular branches of obturator nerve on line H and L were measured by PACS software. Results The proximal motor point (point P) of the gracilis branch, adductor longus branch, adductor brevis branch and adductor magnus branch of the obturator nerve were located at (21.48±1.80)%, (25.85 ± 1.23)%, (28.07 ± 1.65)% and (29.18 ± 2.07)% of the lateral of pubic tubercle along thigh width (line H) respectively; at (8.83 ± 1.01)%, (8.83 ± 1.01)%, (7.57 ± 0.63)% and (7.57 ± 0.63)% of the distal of pubic tubercle along thigh length (line L) respectively. The distal motor points (points D) were located at (16.9 ± 1.33)%, (27.70 ± 2.15)%, (31.18 ± 2.18)% and (35.78 ± 2.79)% of the lateral of pubic tubercle along thigh width (line H) respectively; at (35.57 ± 2.77)%, (26.9 ± 1.96) %, (24.26 ± 1.91)% and (28.04 ± 2.17)% of the distal of pubic tubercle along thigh length (line L) respectively. Conclusion Precise location of these targets can improve the safety and effectiveness for blocking the extramuscular nerve targets in medial femoral muscles spasticity.

    References | Related Articles | Metrics
    Effect of astragalus polysaccharide on histological structure, enzyme activity and Caspase-3 protein expression in the kidney of mice injured by ephedrine
    WEI Ping-ting LI Chong-yang YU Shi-yuan* MENG Ru YUAN Jing
    2015, 46 (4):  564-571.  doi: 10.16098/j.issn.0529-1356.2015.04.021
    Abstract ( )  

    Objective To investigate astragalus polysaccharide(APS)on histological structure and enzyme activity in kidney of mice injured by ephedrine. Methods Seventy-two Kunming mice were divided into 4 groups:the nature control group, the experimental control group,the APS1 and 2 groups. The experimental control group, and APS1 and 2 groups were intraperitoneally injected with 4.0 g/L of ephedrine, 0.2ml twice a day for 15 days. The APS1 and 2 groups were respectively intragastric ally administed with 0.3 ml APS (12.5 g/L,25g/L) 1 hour after ephedrine injection, the control group was injected and intragastric ally administed with the same amount of saline.The blood urea nitrogen (BUN) and creatinine(Cr) content of plasma as well as the activity of superoxide dismutase (SOD) and the malondialdehyde (MDA) content of kidney tissue in the kidney were determined by colorimetry. The kidney histological structures were observed by optical microscopy. The changes of Caspase-3 protein expression in kidney were detected by immunohistochemistry and Western blotting. Results The BUN and Cr content in the blood of the experimental control groups were higher than that of the nature control group. The SOD activity in kidney of the experimental control groups was lower than the nature control groups, while the content of MDA was higher than the nature control groups (P<0.05 or P< 0.01).The histological structure lesions of the experimental control group showed that the glomerular swelled seriously, the renal capsule lumen stenosis,the swelling and necrosis of the renal tubular epithelial cells accured in different degrees. The expressions of Caspase-3 protein in kidney of the experimental control groups were higher than the control groups (P<0.05 or P<0.01). The BUN and Cr content of plasma of the APS groups were lower than the experimental control groups. The SOD activity in the renal tissue of the APS groups increased,while the MDA content decreased (P<0.05 or P<0.01). The histological structure lesions of the APS groups were relieved, the renal capsule lumen were obvious, the swelling of the renal tubular reduced, the degeneration and exfoliating of the epithelial cells alleviated. The expression of Caspase-3 protein in kidney declined significantly (P<0.05 or P<0.01). Conclusion APS enhanced the cells antioxidant enzyme activity, and reduced the expression of Caspase-3 protein.Therefore, APS alleviated the damage of kidney tissue caused by ephedrine effectively, and significantly protected the mice form the ephedrine effects.

    References | Related Articles | Metrics
    A quantitative study of cranial cavity area on mid-sagittal MR images from normal adults
    LIU Xia ZHAO Ji-ping DUAN Chong-feng LI Yang SUI Qing-lan*
    2015, 46 (4):  538-542.  doi: 10.16098/j.issn.0529-1356.2015.04.016
    Abstract ( )  

    Objective To measure the area of the cranial cavity on mid-sagittal magnetic resonance images (MRI) from normal adults, to determine its normal range and to investigate the correlation between the area and age, and its gender difference. Methods A total of 720 normal adults (without diseases associated with the head) were selected stochastically from the PACS of our hospital. The subjects were divided into 6 age groups: 20-29, 30-39, 40-49, 50-59, 60-69, and 70-79 years old. There were 120 subjects per age group with 60 males and 60 females. The areas of the supratentorial, subtentorial, whole cranial cavity and the supratentorial to the subtentorial cranial area ratio of the participant on mid-sagittal MRI were measured. The results were statistically analyzed. Results A slightly positive linear correlation was observed among the areas of the supratentorial, whole cranial cavity and the supratentorial to the subtentorial cranial area ratio and the age. A slightly negative linear correlation was observed between the areas of the subtentorial cranial cavity and the age. The means of the supratentorial, subtentorial, whole cranial cavity and the supratentorial to the subtentorial cranial area ratio were significantly different in different sexes(t=11.55;t=15.69;t=14.70;t=-4.13,all P<0.01). All the measurements was larger in men than in female except for the ratio of the supratentorial to subtentorial cranial areas. Conclusion The reference ranges of all participants in normal adults on mid-sagittal MRI are regular, which can be used in clinic to evaluate the cranial cavity as one indicator.

    References | Related Articles | Metrics
    Identification of hepatocellular carcinoma associated N-linked glycoproteins by two dimensional electrophoresis and mass spectrometry
    WANG Ming-li HAN Da-zheng CHAO Wei-xia LIU Rui-min ZHAO Yun-gang ZHANG Tian QI Yi-jun*
    2015, 46 (4):  495-502.  doi: 10.16098/j.issn.0529-1356.2015.04.009
    Abstract ( )  

    Objective To identify differentially expressed N-linked glycoproteins associated with hepatocellular carcinoma (HCC). Methods Multi-lectin affinity chromatography comprising concanavalin A (ConA), lentil lectin (LCH), snowdrop lectin (GNA) were utilized to isolate N-linked glycoproteins from human immortalized liver cell line L02 and HCC cell lines Huh7, PLC5 and SNU449, followed by 2 dimensional electrophoresis-based quantification and MS/MS identification. Western blotting was used to verify different expression of translationally-controlled tumor protein (TCTP), epithelial cell adhesion molecule (EpCAM) and annexin A2 between two immortalized cell lines vs six HCC cell lines, HCC vs adjacent nontumor liver tissue. Invasion potential in vitro was examined after si-RNA mediated TCTP gene scilencing. Results A total of 42 proteins/isoforms including 14 up-regulated and 28 down-regulated proteins/isoforms were identified. These proteins/isoforms represented 32 unique proteins of which 22 had more than one glycosylation site predicted by NetNGlyc 1.0. These differentially expressed proteins plaid biological functions in redox homeostasis, carbohydrate/energy metabolism, glycolysis, anti-apoptosis, etc. Western blotting validated the up-regulated expression of TCTP, EpCAM and Annexin A2 in 6 HCC cell lines and HCC tissue in comparison with two immortalized cell lines L02, chang liver and adjacent non-tumor tissue, respectively. siRNA mediated downregulation of TCTP remarkably inhibited the invasion potential of in SUN449. Conclusion TCTP, EpCAM and annexin A2 may participate the pathogenesis of HCC and TCTP may become one of molecular targets for the targeted therapy of HCC.

    References | Related Articles | Metrics
    Ultrastructure of lymphatic capillaries with tumor growth in Kunming mice transplanted with S180 sarcoma
    MA Nan ZHANG Li-nan WANG Long-kai ZHAGN Xiao-bo BI Xiu GAO Zi-bin*
    2015, 46 (4):  503-508.  doi: 10.16098/j.issn.0529-1356.2015.04.010
    Abstract ( )  

    Objective To investigate and explore the ultrastructural change of lymphatic capillaries in Kunming mice transplanted with S180 sarcoma with tumor growth. Methods Fifteen Kunming mice were inoculated with S180 ascites via subcutaneous injection to establish the cancer model and divided into five groups. The ultrastructural features of lymphatic capillaries of S180 sarcoma transplanted in Kunming mice were observed by a transmission electron microscope (TEM) in order to analyze the ultrastructural change of lymphatic capillaries with tumor growth. Results The integral lymphatic capillary wall, the anchoring filament and collagen fiber, and the organelles with normal physiological structure were observed in 3 days group. The dissolved or damaged degree of the lymphatic capillary wall and organelles became more and more serious, and the number of the anchoring filament and collagen became less and less with tumor growth in 5 days group, 7 days group and 9 days group. The lymphatic capillary wall was completely destroyed or broken, and any anchoring filament and collagen fiber were scarcely observed, while the lymphatic capillary endothelial cells and the organelles were scarcely observed or identified in 11 day group. Conclusion With the development of transplanted S180, the tumor volume and weight were increased and the dissolved or damaged degree of the lymphatic capillary wall and the organelles became progressively worse.

    References | Related Articles | Metrics
    Establishment of a tumor-bear mouse model carrying human HepG2 cells and exploration of immune tolerance
    LIU Chun-jia ZHANG Xing-yu SHAN Zhi-yan WU Yan-shuang LEI Lei*
    2015, 46 (4):  509-513.  doi: 10.16098/j.issn.0529-1356.2015.04.011
    Abstract ( )  

    Objective To establish anin vivo human hepatoma model carried by mice with normal immune background and to investigate the effect of immune tolerance. Methods We firstly generated green fluorescent protein (GFP) expressed HepG2 cells and intrauterine injected these cells into the peritoneal cavities of E16.5 day mouse fetus. Then we observed the change in newborn mice with bearing cancer and detect the human alpha fetoprotein (AFP) expression in mouse serum by elisa analysis. We subcutaneously injected the HepG2 cells into the adult mice and detected the lymphocyte subgroup ratio of spleen cells by flow cytometry. Results GFP-HepG2 cells were grown in fetus liver and secreted human AFP. After injection of GFP-HepG2 cells in fetus liver, some adult mice developed subcutaneous tumors. Flow cytometry analysis indicated that immune system of tumor-bear mice remained weak attack to human live cancer cells. Conclusion The tumor-bear model carrying human HepG2 cells may be established by injecting the GFP-HepG2 cells into the peritoneal cavities of mouse fetus, parts of which acquire the ability of immune tolerance.

    References | Related Articles | Metrics
    Expression and regulaition of survivin shRNA interference on nuclear factorκB in esophageal ECA109 cells
    LI Xiu-mei CHEN Yan JIANG Xiao-fang LI HuiGUO Qiong LI Hui-wu SHEN Hong-ming*
    2015, 46 (4):  514-520.  doi: 10.16098/j.issn.0529-1356.2015.04.012
    Abstract ( )  

    Objective As the effect of Survivin, a gene expression regulatory factor, on the nuclear factor (NF)-kappa B gene transcription and protein activity remains unknown, this subject aims to explore the possible regulation mechanism of Survivin and NF-kappa B. Methods Survivin shRNA interference technology was applied to liposome transfect esophageal ECA109 cells, detect Survivin silence result, choose the best transfection concentration; Half quantitative RT-PCR was applied to detect genes expression of NF-kappa B and upstream regulatory factors inhibitor of NF-kappa B kinase alpha(IKK alpha), IKK beta in the same pathway; Western blotting was used to detect protein expression of the NF-kappa B. Flow cytometry was used to detect the proliferation and apoptosis of cell lines index changes after ECA109 esophageal cells transfection. Results After survivin shRNA transfection in esophageal ECA109 cells, survivin mRNA and protein expression were obviously down-regulated. NF-kappa B mRNA expression had no obvious difference, however the phosphorylated proteins level of NF-kappaB reduced and its upstream factor IKK alpha, IKK beta mRNA express reduced as well. Esophageal cell ECA109 apoptosis increased significantly, the cells ratio increased in G2 phase but decreased in S phase, and the cell cycle was blocked. Conclusion Survivin regulates NF-kappa B activation by down-regulation of IKK alpha and IKK beta mRNA expression, suggesting that survivin may be a new gene regulatory factor in the tumor signal control network.

    References | Related Articles | Metrics
    Apoptosis inhibitor 5 gene silenced by miRNA-224 in  paclitaxel  inducing HepG2 cells apoptosis
    SHAN Chang-min* SUN Ye-ying LI Juan WANG Dong YAO Qing-shou TAN Xiu-hua LIU Xiang-yong
    2015, 46 (4):  521-527.  doi: 10.16098/j.issn.0529-1356.2015.04.013
    Abstract ( )  

    Objective To explore the gene silencing of microRNAs in paclitaxel inducing HepG2 cells apoptosis.Methods After treated with different concentrations of paclitaxel, apoptosis of human hepotoma cell line HepG2 was detected by flow cytometry, expressions of miR-224 and apoptosis inhibitors 5(api-5) mRNA were determined by Real-time PCR, the expression of API-5 protein was detected by immunohistochemical two steps method and Western blotting method. Results In paclitaxel inducing HepG2 cells apoptosis, the expressions of miR-224 in HepG2 treated with 0.5 mg/L, 1mg/L paclitaxel for 24hours decreased slightly, while those of the others increased obviously by analysis of 2-ΔΔCT method. The expressions of api-5 mRNA in HepG2 treated with 0.5 mg/L, 1mg/L paclitaxel for 24hours decreased slightly, while those of the others were up-regulated except that those of treatment with 0.5mg/L paclitaxel and the control was similar. By the treatment of Image Pro Plus v 5.1,it was found that the levels of API-5 protein were down-regulated with treatment of paclitaxel through the analysis of t-test(P<0.05). Conclusion Increased miR-224 may affect the expression of its target gene api-5. Through the combination with api-5 mRNA 3’UTR,the completion of translation is prevented, which may impact on liver cancer cell apoptosis pathway.

    References | Related Articles | Metrics
    Expression of annexin A7 in human gastric carcinomas and the clinic significance
    HANG Yang XU Qian CHEN Long WANG Xiao-jie TIAN Huan-na LI Chun-hui LI Xin*
    2015, 46 (4):  528-531.  doi: 10.16098/j.issn.0529-1356.2015.04.014
    Abstract ( )  

    Objective To dectect the expression of annexin A7 in human gastric carcinoma so as to detect the mechanism. Methods Fifty five gastric carcinoma samples and 25 normal human gastric samples were collected. Each sample was lysed in lysis buffer. Whole protein of the supernatant was estimated by BCA-100 Protein Quantitative Analysis Kit. The expression of annexin A7 in human gastric carcinoma and normal gastric tissue was examined with Western blotting and Real\|time PCR. Immunehistolchemistry was used to detect the expression position of annexin A7 in the gastric carcinoma cells. Results The expression of annexin A7 increased in gastric cancer not only on protein level but also on RNA level when compared with normal gastric tissues, the difference was significant (P<0.05); Annexin A7 mainly expressed in the cytoplasm of the gastric carcinoma cells. Conclusion Expression of annexin A7 is up-regulated in human gastric cancer, and it may involve in the development of gastric cancer.

    References | Related Articles | Metrics
    Olive leaf extract improves cognition and reduces oxidative damage and apoptosis in mice with heroin dependence
    WANG Yu* WANG Sheng-qing CUI Wen-hui QI Lin-lin
    2015, 46 (4):  462-470.  doi: 10.16098/j.issn.0529-1356.2015.04.005
    Abstract ( )  

    Objective To observe the effect of olive leaves extract (OLE) on cognition, oxidative damage and hippocampal apoptosis in heroin-dependent mice. Methods One hundred and fifty mice were given subcutaneous administration of heroin (10 mg/ kg) for 40 days, and then treated with different doses (0, 50, 100, 200 mg/kg) of OLE. The spatial learning and memory abilities were evaluated with Morris water maze test. Changes of the cell structure of hippocampus were observed by transmission electron microscopy. Spectrophotometry was used to detect the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione (GSH) and malondialdehyde (MDA) content. Bax protein and caspase-3 expression in the hippocampus were determined by immunohistochemical and Western blotting analysis. Results Hippocampal neurons in the model group showed irregularity of nuclear membrane, abnormal or vacuolar structures of organelles. The escape latency and escape distances in the model group were significantly longer and the time spent and swimming distance percentage in the target quadrant were significantly shorter than that in the normal control group (P<0.01). The activities of SOD, CAT, GPx, and GSH in the model group lower than those of the normal control group, while the content of MDA was higher than that of normal control group (P<0.01). The expressions of Bax protein and Caspase-3 in the hippocampus of the model group were increased compared with the normal control group (P<0.01). Administration of OLE alleviated neuronal injuries and reduced damage to organells, and shortened the escape latency and escape distances, prolonged the time spent and swimming distance percentage in the target quadrant. Moreover, the activities of SOD, CAT, GPx, and GSH in OLEtreated groups increased, while the MDA content and expressions of Bax protein and Caspase-3 decreased, in a dose-dependent manner (P<0.05 or P<0.01). Conclusion The neuroprotective effects of OLE on heroininduced brain damage are associated with its properties of antioxidant and inhibiting apoptosis.

    References | Related Articles | Metrics
    Expression of gene associated with retinoic-interferon-induced mortality-19 and its effect on mouse embryo implantation
    SUN Yan-yan DENG Xiao-hui CHAO Lan* YANG Yang CHENG Lai-yang DU Tiao-qi WANG Jing
    2015, 46 (4):  471-479.  doi: 10.16098/j.issn.0529-1356.2015.04.006
    Abstract ( )  

    Objective To explore the expression of gene associated with retinoic-interferon-induced mortality-19(GRIM-19) gene and the effect of GRLM-19 on the embryonic implantation. Methods The expression of GRIM-19 in the pregnant mouse uterus was detected by immunohistochemistry. The expression of GRIM-19 in the mouse uterus was detected by Real-time PCR and Western blotting with early pregnancy,hormone treatment and pseudopregnancy models. TUNEL method was used to detect apoptosis in uterine tissues of pregnant mice. An optimized model of R95-2 spheroids and a Bewo monolayer co-culture model were employed to study the effect of GRIM-19 in the embryo-endometrial epithelium attachment. Apoptosis was assayed by flow cytometry. Mitochondrial membrane potential was assayed by JC-1. Western blotting and Real-time PCR were used to detect signal transducer and activator of transcription(STAT3), tumor necrosis factor(TNF)-α and interleukin (IL)-11 mRNA and protein level. Results The immunohistochemical positive staining for GRIM-19 was distributed in the luminal epithelium and glandular epithelium in pregnant mice. The intensities of both GRIM-19 protein and mRNA positive staining were decreased to the lowest level at pregnancy day 4 in the pregnancy group, no significant difference in the pseudopregnancy group, and lower in the estrogen treatment group. Overexpression of GRIM-19 made RL95-2-BeWo co-culture spheroid adhesion rate decreased, apoptosis increased, and mitochondrial membrane potential reduced; STAT3 and IL-11 mRNA and protein levels reduced, but TNF-α protein and mRNA levels increased. Conclusion GRIM-19 plays an important role in embryo implantation, possibly through the regulation of apoptosis and immune tolerance to provide protection for the embryo implantation.

    References | Related Articles | Metrics
    Synthesis and toxicity study on the single- and double-stranded aspartic acid and lysine peptide
    ZHAO Qian-yi XU Ting LI Meng-hua YAN Wen-juan WANG Shu-pei XU Cun-shuan*
    2015, 46 (4):  480-487.  doi: 10.16098/j.issn.0529-1356.2015.04.007
    Abstract ( )  

    Objective To understand the toxicity and application prospect of polypeptide formed by the same amino acids. Methods This study focused on the solid-phase polypeptide synthesis of Asp and Lys polypeptide, and analyzed their toxicity by the method of tail vein injection to observe the lethal effect on mice. Results When the concentration of the nine kinds of single-strand asp-peptides were 0.15, 0.1, 0.09, 0.03, 0.0065, 0.03, 0.034, 0.035and 0.04 mol/L, respectively, the death rate of the BALB/c mouse was LD50. When the concentration of the nine kinds of single-strand lys-peptides were 0.28, 0.1, 0.047, 0.0225, 0.0028, 0.00166, 0.0015, 0.0011and 0.00075 mol/L, respectively, the death rate of the BALB/c mouse was LD50. When the concentration of the ten kinds of double-strand asp-lys-peptides were 0.5, 0.05, 0.017, 0.014, 0.009, 0.006, 0.004, 0.004, 0.004 and 0.004 mol /L, respectively, the death rate of the BALB/c mouse was LD50. Conclusion The toxicity of single-strand peptides was greater than that of double-strands. With the number of amino acids increasing, the toxicity of the single- strand asp-polypeptide, from dipeptide to hexapeptide, was increased, while Asp-heptapeptide to Asp-decapeptide was decreased. In the case of Lys-polypeptide, the toxicity of the single-strands was increased when the number of amino acid was increased. For the double-stranded Asp-Lys- polypeptide, by increasing the number of amino acid, the toxicity of the dipeptide to hexapeptide was increased, while heptapeptide to decapeptide had no change.

    References | Related Articles | Metrics
    Expression of nanog protein in fibroblasts from multiple organs of the neonatal rat
    LU Zhao-hui CHANG Yu-qiao GUO Zhi-kun* ZHAO Guo-an LI Qiong LI Ci-xia
    2015, 46 (4):  488-494.  doi: 10.16098/j.issn.0529-1356.2015.04.008
    Abstract ( )  

    Objective To investigate different expression and its significance of nanog protein in multiple organs fibroblasts of neonatal rats, including heart, liver, spleen, lung, kidney and skin. Methods The expresssion and co-expression of nanog and vimentin in fibroblasts were detected in multiple organs by immunohistochemistry and immunofluorescence. Fibroblasts in multiple organs cultured in vitrowere labeled with nanog and vimentin by immunofluorescence. Results Nanog and vimentin were detected in six organs in neonatal rats respectively. Co-expression rates of these two proteins were(10.07±0.77)% in heart,(9.97±3.70)% in liver,(11.23±2.98)% in spleen,(15.07±2.87)% in lung,(11.75±2.76)% in kidney and(16.14±1.28)% in skin. So co-expression rates of skin and lung were higher than that of other four organs(P<0.05). Conclusion Fibroblasts in multiple organs which can express nanog may have stem cell-like characters and fibroblasts in skin and lung of neonatal rats have more potential for organ repair.

    References | Related Articles | Metrics