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    2014, Volume 45 Issue 4
    06 August 2014
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    Dynamic expression of Lhx8 in nerve growth factor induced hippocampal neuroregeneration
    LI Hao-ming ZHU Pei-pei JIN Guo-hua* SHI Jin-hong ZOU Lin-qing TIAN Mei-ling YI Xin QIN Jian-bing
    2014, 45 (4):  441-445.  doi: 10.3969/j.issn.0529-1356.2014.04.001
    Abstract ( )  

    Objective To investigate the relationship between the nerve growth factor (NGF) induced hippocampal neuroregeneration and homeobox gene Lhx8.Methods Seventy-two SD rats were divided into control group, transected group, NGF group, transected combined with NGF group after right fimbria-fornix transection and NGF intracerebroventricular injection. Real-time PCR and Western blotting were applied to detect the gene and protein expression of Lhx8 in each group. The choline acetyltransferase (ChAT)/Lhx8 double labeled cells in subgranular zone (SGZ) of hippocampus in each group were detected by immunofluorescence. Results The expression of Lhx8 gene and protein in the transected, NGF group and especially in the transected combined with NGF group was obviously higher than in the control group. The number of ChAT/Lhx8 double labeled cells in the NGF group and the transected combined with NGF group was obviously more than in the control group and transected group. Conclusion The hippocampal neuroregeneration which induced by NGF intracerebroventricular injection was associated with the higher expression of Lhx8.

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    Low dose of genistein attenuates neuronal injury and improves learning and memory functions of rats following global cerebral ischemia
    MA Wen-dong TU Jing-yi ZHU Ying ZHANG Xi TANG Hui WANG Rui-min*
    2014, 45 (4):  446-451.  doi: 10.3969/j.issn.0529-1356.2014.04.002
    Abstract ( )  

    Objective To explore the neuroprotective role of Genistein (GEN) on hippocampal CA1 neurons and the possible mechanism following global cerebral ischemia (GCI) in rats. Methods Seventy five rats were subjected to global cerebral ischemia (GCI) by four-vessel occlusion and randomly divided into five groups, sham, ischemia/reperfusion (I/R), GEN, ICI 182,780 and vehicle groups. Fluoro-Jade B and neuron-specific nuclear-binding protein (NeuN) staining was used to observe CA1 neuronal survival. TUNEL was used to detect apoptotic neurons. Spatial learning and memory function of the rats were evaluated by Morris water maze. Results The best dose of neuroprotective role of GEN was 1.0mg/kg body weight. Compared with sham, TUNEL-positive neurons in the hippocampal CA1 region increased significantly in I/R and vehicle groups (P<0.01), while posttreatment with GEN (1.0mg/kg) at 5min after ischemia by tail vein injection decreased markedly (P<0.01). Treatment of 1.0mg/kg GEN markedly attenuated spatial learning and memory deficits of the rats after ischemic insult compared to I/R group. Furthermore, ICI 182,780 significantly abolished the neuroprotective role of GEN (P<0.01). Conclusion The low-dose (1.0mg/kg) GEN significantly attenuates neuronal damage and cognitive deficits following GCI in rats, and the mechanism may be involved in estrogen receptor activity.

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    Expression of Caspase-12 in cellular apoptosis of hippocampus in the rat model of posttraumatic stress disorder
    LIU Hong HAN Fang SHI Yu-xiu*
    2014, 45 (4):  452-456.  doi: 10.3969/j.issn.0529-1356.2014.04.003
    Abstract ( )  

    Objective To observe changes of apoptosis and Caspase-12 expression in hippocampus of the rat model of posttraumatic stress disorder (PTSD).Methods Single prolonged stress (SPS) was used to duplicate a rat model of PTSD.Male Wistar rats were randomly divided into 1, 4, 7 days groups after exposure to SPS and a normal control group. Transmission electron microscopy was used to observe morphologic changes of the hippocampus neurons. The expression of Caspase-12 was detected by immunohistochemistry, Western blotting and RT-PCR. Results After SPS, the hippocampus neurons had characteristic morphologic changes of apoptosis, and the expression of Caspase-12 reached the peak at SPS 4d.Conclusion PTSD activates the ERS-mediated apoptosis in hippocampus via Caspase-12, which may be a pathophysiology base of the atrophy of the hippocampus in PTSD.

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    Development of the mouse spinal cord and neuroapoptosis
    DENG Juan ZHENG Hong LI Xue XUE Shuai LI Li-li NIE Meng-yue WU Ping* DENG Jin-bo*
    2014, 45 (4):  457-464.  doi: 10.3969/j.issn.0529-1356.2014.04.004
    Abstract ( )  

    Objective To investigate the neural proliferation, differentiation and apoptosis of the developing spinal cord of the mouse and to discuss the mechanism of spinal cord’s development. Methods 5-Bromodeoxyuridine (BrdU) assay was used to mark the proliferative neural stem cells, and the immunofluorescent stainings (DCX, NeuN and Caspase8) were carried out to visualize the newborn neurons, mature cells and apoptotic cells in the spinal cord with 173 mice arrange from E18 to P90.
    Results BrdU positive neural stem cells appeared evenly in the spinal cord at early days. With age increasing, the neural stem cells differentiated into neuroglial cells and neurons. The newborn neurons in the subventricular zone migrated toward the intermediate zone (putative gray matter) and differentiated into mature neurons gradually. With neurons’ concentrating towards the center, the gray matter formed an “H” shape. In the meantime, with neural differentiation, some apoptotic neurons appeared among the newborn neurons and mature neurons. Double immunostaining showed that most apoptotic neurons were newborn neurons, suggesting the neuroapoptosis more likely occurred in newborn neurons. The statistical data showed that the number of DCX, NeuN and Caspase-8 positive cells reduced with age increasing, suggesting neural differentiation and neuroapoptosis decreased during spinal cord’s development. Conclusion Neural proliferation, neural differentiation and neuroapoptosis occur in developing spinal cord. They work together to regulate the formation and development of the spinal cord.

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    Expression of progesterone receptor in the cervicothoracic ganglion of female goat
    GUO Xiao LI Qiang CHEN Weng-dong XU Yong-ping* JIN Xiu-fang WANG Zhi-hao DONG Wei FAN Jie
    2014, 45 (4):  465-468.  doi: 10.3969/j.issn.0529-1356.2014.04.005
    Abstract ( )  

    Objective  In order to explore whether neurons in the cervicothoracic ganglion of female goat were equipped with the condition for the role of progesterone.  Methods Immunohistochemical SP method was used to detect the expression of progesteron receptor(PR) in cervicothoracic ganglion of female goats. Results The result indicated that PR-immunoreactive-products were mainly present in the cell bodies of neurons,and satellite cells, Schwann cells, neurapophysis and nerve fiber also had weaker staining.The cell membrane and cytoplasm of neurons represented brown as strong positive,but the nucleus of neurons showed the presence of heterogeneity, 85.10% neurons’ nucleus was strong positive,of which 31.91% nucleolus being clear vacuoles were negative; 14.90% nucleus displaying vacuolization had no shaining, of which 7.45% nucleolus was strong positive. There were weak positive and flaxen PR-immunoreactive-products in satellite cells, Schwann cells, neurapophysis and nerve fiber. Image analysis shows that PR of neurons was extremely significant compared with other nonnerve cells(P<0.01). Conclusion The result proved that the sympathetic postganglionic neurons in cervicothoracic ganglion of female goat were the one of main target cells of progesterone,suggesting that progesterone may be involving in the neuroregulation of cardiac functional activities through affectinng the activities of neurons in the cervicothoracic ganglion of female goat,and PR in the cervicothoracic ganglion may act as a network node to coordinate endocrine regulation of progesterone and neuroregulation of autonomic nerve on the cardiac functional activities.

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    Effects of basic fibroblast growth factor on endothelial function and structure of the basilar artery of atherosclerotic rats
    ZHU Jun-de* YU Yan GE Guo
    2014, 45 (4):  469-474.  doi: 10.3969/j.issn.0529-1356.2014.04.006
    Abstract ( )  

    Objective To study the protective effect of basic fibroblast growth factor (bFGF) on endothelial function and structure of the basilar artery of atherosclerotic rats. Methods A total of forty-eight male adult Wister rats were randomly divided into the normal control, the atherosclerosis (AS) model and the bFGF treatment groups. The AS model group and the bFGF treatment group were injected with a single dose of vitamin D3 (6×105IU/kg) and loaded with high fat diet for six consecutive weeks. The bFGF (9.5μg/kg, twice one day) was injection into the abdominal cavity after six weeks in the bFGF treatment group for two weeks, and an identical volume saline was given for the AS model group and the normal control group. After eight weeks, all the rats were sacrificed. The relaxation percentages of the isolated basilar artery in response to acetylcholine (Ach) were detected and the pathological lesions of them were observed under a light microscope. ELISA and colorimetry assayed the content of serum VEGF and basilar arterial nitric oxide (NO). The basilar artery was used for primary culture of both vascular endothelial cells (VECs) and vascular smooth muscle cells (VSMCs). The influence of bFGF on the proliferation vitality of VECs was measured in vitro with MTT assay. TRITC-phalloidin labeling the cytoskeleton microfilament of VSMCs was observed by laser confocal microscopy. Results The early AS plaques were presented after six weeks by hyper lipid foods. Compared with the AS model group, the relaxation percentage of the isolated basilar artery, the content of both serum VEGF and basilar arterial NO in the bFGF treatment group were obviously increased, but the pathologic injury of the basilar artery was significantly decreased (P<0.05). The proliferation vitality of VECs was obviously increased (P<0.05); the cytoskeleton microfilament of VSMCs was of obviously improvement. Conclusion AS may aggravate the basilar arterial injury, but bFGF may efficiently improve the arterial endothelial function and decrease the pathological lesion of the basilar artery in the AS model rats, which may promote the arterial protective effect.

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    Location of trefoil factor 3 in chromophil cells of rat adenohypophysis
    ZHANG Wen-jing SUN Lin WANG Jing ZHANG Jing WU Jing-fang*
    2014, 45 (4):  475-479.  doi: 10.3969/j.issn.0529-1356.2014.04.007
    Abstract ( )  

    Objective To investigate the distribution and expression of trefoil factor 3(TFF3) in the eosinophilic cells and basophilic cells in the pars distalis of the rat pituitary. Methods The immunohistochemiscal staining technique was used to show the coexpression of TFF3/ growth hormone (GH),TFF3/ prolactin (PRL),TFF3/ thyrotroph (TSH),TFF3/ adrenocorticotropin (ACTH),TFF3/ follicle stimulating hormone (FSH),TFF3/ luteinizing hormone (LH) in two contiguous slices. Results The immunoreactive products of TFF3 and chromophil cells were brown granules,mainly expressed in cytoplasm.ACTH positive signals were expressed in the cell membrane and mainly located in the pars distalis hypophyseos.TFF3 existed in parts of GH,PRL,TSH,ACTH,FSH and LH cells in contiguous slices, accounting for 19.4%, 22.4%, 9.2%, 6.5%, 35.7%, 8.3%, respectively, in which FSH was the most, PRL and GH were less. Conclusion TFF3 expresses in GH,PRL,TSH,ACTH,FSH, and LH cells in the pars distalis,which enriches the morphological data for the location of TFF3 in gland cells of pars distalis hypophyseos.

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    Effect of uncoupling protein 2 on activation of hepatic stellate cell in liver fibrosis of rats
    AN Jian-duo JIANG Ying BAI Yun-fei WANG Xue-jiang*
    2014, 45 (4):  480-484.  doi: 10.3969/j.issn.0529-1356.2014.04.008
    Abstract ( )  

    Objective To explore the role of uncoupling protein 2 (UCP2) in the development of hepatic fibrosis and its molecular mechanism. Methods The CCl4-induced liver fibrosis rat modelin vivo was established to observe the pathological changes of rat livers. The expression levels of UCP2 and p38 mitogen activated protein kinase (p38 MAPK) were detected by using the techniques of Western blotting, Real-time PCR and immunohistochemistry. The hepatic stellate cells (HSC) were stimulated by CCl4 and UCP2-specific inhibitor Genipin to mimic liver fibrosis in vitro. The expression levels of UCP2 and p38MAPK were determined by using Western blotting. Results We found that UCP2 and α-SMA expression levels increased significantly (P<0.05, n=10) in the liver of rats with CCl4-induced liver fibrosis when compared with that of the normal control rats in vivo. Similarly, the expression levels of UCP2 and p38 MAPK were up regulated (P<0.05, n=6) in CCl4-treated HSC cells in vitro. However, the expressions of UCP2 and p38 MAPK were down regulated (P<0.05, n=6) in genipin-treated HSC cells in vitro. Conclusion UCP2 is involved in liver fibrosis, and probably contributed to the activation and proliferation of hepatic stellate cells.

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    Effects of transcription factor cAMP response element binding protein on taxolinduced HeLa cell-cycle arrest
    HUANG Shuai-shuai WANG Xue ZHUANG Hai-hui WANG Yu-duo ZHOU Xi-wu WANG Ping*
    2014, 45 (4):  485-492.  doi: 10.3969/j.issn.0529-1356.2014.04.009
    Abstract ( )  

    Objective To explore the effects of cAMP response element binding protein (CREB) on taxol-induced cell cycle arrest in HeLa cells. Methods MTT assay was used to determine the optimal concentration and treatment time. PCR method was used to construct the recombinant plasmid pCI neo/CREB(PN)and site-directed mutagenesis recombinant plasmid pCI neo/CREB-M(PM). Cell cycle was assayed by flow cytometry. Expressions of pCREB, CREB, cyclins and CDKs were assayed by Western blotting. Results The effective conditions of taxol treatment on HeLa cells were 0.1μmol/L for 24 hours. After cells were treated with 0.1μmol/L taxol, G2/M phase was arrested in a time-dependent manner, accomplished with the decrease of cyclin A, a significant increase of cyclin B1, D1 and phosphorylated CREB(pCREB) protein expression, whereas, no marked changes were observed in cyclin E, CDK1, CDK2, CDK4 and CREB expressions. However, combinantion of PM and taxol treatment significantly reduced taxol-induced G2/M phase arrest, and reversed the effect of taxol-decreased cyclin A, increased cyclin B1 and D1 expression. Conclusion Tanscription factor CREB-mediated specific cyclins play a pivotal role in taxol-induced G2/M arrest in HeLa cells.

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    Proteomic fingerprinting of N-linked glycoproteins involved in hepatocellular carcinoma
    MA Jin QI Yi-jun* LIU Rui-min WANG Ming ZHANG Tian ZHU Han MA Yuan-fang
    2014, 45 (4):  493-499.  doi: 10.3969/j.issn.0529-1356.2014.04.010
    Abstract ( )  

    Objective To identify differentially expressed N-linked glycoproteins between hepatocellular carcinoma (HCC) and adjacent non-tumorous liver tissues.
    Methods N-linked glycoproteome was extracted by multi-lectin affinity chromatography comprising concanavalin A (ConA), lentil lectin (LCH), and snowdrop lectin (GNA) and subsequently subjected to two-dimensional electrophoresis (2DE) and mass spectrometry(MS) for identification of differential glycoproteins between 10 pairs of HCC and adjacent non-cancer tissue. Western blotting was used to verify different expression of human liver carboxylesterase1 (hCE1), haptoglobin (HP)and cathepsin D (CD). Invasion potentialin vitro was examined after si-RNA mediated CD gene scilencing. Results LC-ESI-MS/MS identified a total of 28 differentially expressed glycoproteins (14 up-regulation and 14 down-regulated). Western blotting detected consistent down-regulation of hCE1 and HP, and up-regulation of pro-cathepsin D (pCD) in HCC. Up-regulation of ConA-binding CD (ConA-CD), however, was verified in HCC only after ConA-CD enrichment by ConA chromatography. Down-regulation of CD expression mediated by CD-siRNA markedly inhibited the in vitroinvasive potential of SNU449 and SNU473.
    Conclusion Dysregulation of HP, hCE1 expression and alteration of glycans linked to CD may play crucial roles in pathogenesis of HCC.

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    Effects and mechanisms of periostin overexpression on invasion and migration of the nasopharyngeal carcinoma 6-10B cell line
    WANG Hui-jie SHI Jin-feng XIE Yuan-jie ZENG Gu-qing DU Ya-lan HUANG Xing-qiong LONG Zhi-feng YU Jiang-dong LI Mei-xiang*
    2014, 45 (4):  500-506.  doi: 10.3969/j.issn.0529-1356.2014.04.011
    Abstract ( )  

     Objective To explore the effects and mechanisms of periostin overexpression on migration and invasion of nasopharyngeal carcinoma(NPC) cell line. Methods The recombinant plasmids [pCMV-neo (+)-periostin] and control plasmids [pCMV-neo (+)] were transfected into 6-10B cells using lipofectamine 2000TM reagent. The expression of periostin was detected with PCR and Western blotting. Transwell chamber invasion assay was employed to assay the migration and invasion of 6-10B cells before and after transfection. A gelatin zymogram was used to detect the activity of MMP-2 and MMP-9 in cultivated supernatant of 6-10B cells before and after transfection. The expression of integrin-αvβ5 was detected by immunohistochemistry(IHC) in 6-10Bperiostin cells, 6-10Bvector cells and 6-10B cells as well as normal nasopharyngeal mucosa (NNM) and NPC and at the same time periostin also was detected by immumohistochemistry in NNM and NPC, and densitometry analysis using image-pro plus 6.0 software, and the correlation between periostin and integrin-αvβ5 on NPC was assayed with statistics. Results Over expression of periostin promoted cell migration and invasion. The expression levels of integrin-αvβ5 in primary NPC and 6-10Bperiostin cells were significantly higher than those in NNM and 6-10Bvector, 6-10B cells. The expression in NPC of integrin-αvβ5 showed positively correlated with the expression of periostin (r=0.682, P<0.01). Conclusion Periostin plays an important role in regulation of cell migration and invasion probably by combining with integrin-αvβ5 to improve the activities of MMPs.

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    Comparative proteomic analyses on the differentiation of dimethyl sulfoxide induced murine erythroleukemia cells
    YANG Zu-li ZHU Xiao-fang SHI Ming-ming HU Fan ZHAO Fu-kun ZHANG Shi-fu*
    2014, 45 (4):  507-515.  doi: 10.3969/j.issn.0529-1356.2014.04.012
    Abstract ( )  

    Objective To explore the differentially expressed proteins during erythroid differentiation. Methods The murine erythroleukemia (MEL) cell were treated by DMSO, and the comparative proteomic was systematically analyzed and identified on different differentiating time points. ratio of cell differentiation and viability were detected by benzidine staining, MTT assay and Ter119 immunofluorescence. Using two-dimensional gel electrophoresis combined with mass spectrometry technology and bioinformatics analysis, we conducted a comparative proteomic analysis on MEL cells during the process of induced differentiation to screen and identify differential proteins. Results The MEL cells induced by 1.2% DMSO for 0 hour, 6hours, 12hours, 24hours, 36hours, 48hours, 72 hours, 96 hours, 120 hours were collected for proteomic analysis, by two-dimensional gel electrophoresis combined with mass spectrometry. A total of 87 kinds of proteins were successfully identified. MEL cells exposed to DMSO at a final concentration of 1.2% for 120 hours reached the highest differentiation rate of 67%. MTT assay showed that 1.0%, 1.2%, 1.4% DMSO had no inhibiting effect on cell vitality. Conclusion DMSO may induce MEL cells to differentiate and have no inhibiting effect on cell vitality. The 87 kinds of differentially expressed proteins from two-dimentional gel electrophoresis may be divided into twelve categories; the most three parts are 41% enzyme protein, 15% structural protein and 13% regulatory protein.

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    Expression of NonO protein during murine erythroleukemia cell differentiation induced by sodium butyrate
    SANG Ting-ting HU Jiang-jiang XUE Jian-you QI Wu-lin ZHAO Fu-kun ZHANG Shi-fu*
    2014, 45 (4):  516-520.  doi: 10.3969/j.issn.0529-1356.2014.04.013
    Abstract ( )  

    Objective To study NonO protein expression changes in murine erythroleukemia (MEL) cell differentiation induced by sodium butyrate.Methods Benzidine staining was used to test sodium butyrate-induced erythroid differentiation of MEL cells. We detected NonO protein expression changes in MEL cell differentiation induced by sodium butyrate and NonO protein localization in MEL cells by Western blotting and immunocytochemistry. Furthermore, we applied PCR technique to detect NonO RNA expression in differentiation process. Results We found that NonO protein was upregulated at gene and protein levels in the erythroid differentiation process of MEL cells induced by sodium butyrate and located in cytoplasm and nucleus in MEL cells. Conclusion These results show that NonO protein is closely related with MEL cell differentiation induced by sodium butyrate,which may provide important clues for further study of the mechanism of leukemia.

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    5-Aza-2’-deoxycytidine and 4-phenylbutyric acid exert a cooperative effect on the expression of miR-196b in chronic myeloid leukemia cells
    LIU Yue SHUAI Chun LI Jie-sheng YIN Hong SONG Yan-bin MA Wen-li*
    2014, 45 (4):  521-524.  doi: 10.3969/j.issn.0529-1356.2014.04.014
    Abstract ( )  

    Objective To study if 5-Aza-2’-deoxycytidine along or together with 4-phenylbutyric acid could affect miR-196b expression levels in chronic myeloid leukemia cells.
    Methods K562 cells were treated with DNA methylation inhibitor 5-Aza-2’-deoxycytidine, histone deacetylase inhibitors 4-phenylbutyric acid separately and the combined treatment with both of them, then expression levels of miR-196b were detected using Real-time PCR. Results The half inhibition concentration of 4-phenylbutyric acid was 1.58mmol/L. Comparing with the expression level of miR-196b in normal human bone marrow cells, the expression levels of miR-196b were significantly lower in Aza group, PBA group and negative control cells and nearly consistent among three groups, and as high as normal cells in combined treatment group. Conclusion The expression level of miR-196b in K562 cells could not return to normal treated with 5-Aza-2’-deoxycytidine or 4-phenylbutyric acid separately, while could restore normal when treated with both agents, indicating that miR-196b expression level in K562 cells is related with both DNA methylation and histone acetylation.

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    Effects of ganoderma lucidum spore oil and ganoderma lucidum extraction spore oil on angiogenesis regulatory factors
    ZHANG Jing NIU Miao-miao YANG Li FAN Li-sha WU Li ZHAN Jun* ZHANG Hong-quan
    2014, 45 (4):  525-530.  doi: 10.3969/j.issn.0529-1356.2014.04.015
    Abstract ( )  

    Objective To study the role of ganoderma lucidum spore oil and ganoderma lucidum extraction spore oil in neovascularization of human high malignant breast cancer. Methods Human high malignant breast cancer cell MDA-MB-231 and tumor-bearing nude mice established with MDA-MB-231 were treated with different doses of ganoderma lucidum spore oil and ganoderma lucidum extraction spore oil. Epidermal growth factor receptor(EGFR) expression level was examined by Western blotting and the RNA expression levels of neovascularization related molecules such as EGFR, vascular endothelial growth factor(VEGF), metalloproteinases(MMPs), thrombospondin(TSP-1), platelet derived growth factor(PDGF), fibroblast growth factor(FGF) were detected by Real-time PCR. Results Both ganoderma lucidum spore oil and ganoderma lucidum extraction spore oil inhibited the expression of EGFR in vitroand in vivoin a dose-dependent way. Both compounds induced down-regulation of VEGF and up-regulation of TSP-1 at RNA level. The effect of Ganoderma lucidum extraction spore oil was more significant than that of ganoderma lucidum spore oil. Conclusion Both ganoderma lucidum spore oil and ganoderma lucidum extraction spore oil inhibite the expression of neovascularization related molecules and increase the expression of molecules inhibiting neovascularization, whereas the effect of ganoderma lucidum extraction spore oil is more obvious.

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    Anatomy of the superficial epigastric artery perforator flap in rats
    FANG Dong LIN Hai-shuang YU Feng ZHANG Si-fang DING Mao-chao CUI Huai-rui* TANG Mao-lin XU Da-chuan
    2014, 45 (4):  531-535.  doi: 10.3969/j.issn.0529-1356.2014.04.016
    Abstract ( )  

    Objective To investigate the anatomy of the superficial epigastric artery perforator flap, and to provide anatomical basis for harvesting flap. Methods Of 27 SD rats, 7 were used for gross anatomy observation and anatomic characteristics and 20 rats for lead oxide-gelatin injection followed by computer picture processing, measurements and the related parameters recording. Results The superficial epigastric artery originated from femoral artery, and gave off its first branch when passed through the superficial fascia. The trunk branched into a lateral perforator and a medial perforator, which anastomosed with thoracodorsal artery and lateral thoracic artery, respectively. The average external diameter of superficial epigastric artery was(0.46±0.02)mm at its starting point,and(0.46±002)mm at the superficial fascia level. The nutritive area of superficial epigastric artery was (18.37±3.67) cm2. The anastomosed area with thoracodorsal artery and lateral thoracic artery was(5.34±0.86)cm and(6.28±0.29)cm, respectively, away from the horizontal line through axillary,and (4.38±0.38)cm and(2.04±0.33)cm, respectively, away from the ventral median line. Conclusion The position and external diameter of superficial epigastric artery are constant, and the superficial epigastric artery perforator flap is a ideal flap model for research on free flap transplantation, flap supercharging, and hemodynamics.

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    Digital 3D anatomical modeling of the mandible with full teeth
    ZHU Fu-dong SHI Jue SONG En GE Wei-li LIAO Sheng-hui*
    2014, 45 (4):  536-539.  doi: 10.3969/j.issn.0529-1356.2014.04.017
    Abstract ( )  

    Objective To study and establish a high quality digital 3D anatomical modeling of the mandible with full teeth.Methods A set of accurate digital models of standard anatomical specimens of mandibular teeth were obtained by laser scanning, and the 3D mandible model was reconstructed by CT scan data; then, a registration deformation method based on the geometry and image anatomical landmark was employed to do the registration of each tooth to the mandible model, and finally the tooth enamel, dentin, periodontal ligament were generated. Results A high quality digital 3D anatomical modeling of the mandible with full teeth was built, each tooth had detail crown and whole root, the distinction between the enamel, dentin, periodontal ligament, and any anatomical regions can be zoomed and rotately displayed.
    Conclusion The digital 3D anatomical modeling of the mandible with full teeth has realistic 3D imaging view and convenient teachinglearning function, and has tremendous apllication futures in the stomatology, maxillofacial and other medical departments.

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    Spatio-temporal expression of P2X3 receptor in rats with diabetic mechanical allodynia
    CUI Yuan-yuan WU Huang-hui WANG Lan SHI Juan LI Yun-qing*
    2014, 45 (4):  540-544.  doi: 10.3969/j.issn.0529-1356.2014.04.018
    Abstract ( )  

    Objectiv To investigate the spatio-temporal expression of P2X3 receptor (P2X3R) in rats with diabetic mechanical allodynia (DMA).Methods DMA model in rats was induced by intraperitoneal injection of streptozocin (STZ). The von Frey filaments were applied to identify the changes of the paw withdrawal threshold (PWT) in DMA rats. Immunofluorescence was employed to detect the spatiotemporal expression of P2X3R in the spinal dorsal horn (SDH), dorsal root ganglion (DRG) and hind paw skin on different time points after intraperitoneal injection of STZ, respectively. The protein expression of P2X3R in SDH and DRG was further semi-quantitatively analyzed by Western blotting. Results Compared with control group, intraperitoneal injection STZ induced significant mechanical allodynia indicated by the reduced PWT from 7 days, and which reached the peak on 14d and maintained to 28days (P<0.05). The expression of P2X3R in DRG neurons was significantly increased on 14 days and 21 days (P<0.05), while that in SDH and skin was markedly increased on 21 days and 28 days, compared with control group (P<0.05). Conclusion With the progress of DMA, the expression of P2X3R was significantly increased in the SDH, DRG and skin, which was almost parallel with the mechanical allodynia, but the changes in SDH and skin were 1 week later than that in DRG. These results suggest that P2X3R may play a key role in the maintenance of the DMA.

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    Expression of neurofilament protein and synaptophysin in the developing spinal cord of human embryo
    ZHANG Yong LIU Xue-hong*
    2014, 45 (4):  545-549.  doi: 10.3969/j.issn.0529-1356.2014.04.019
    Abstract ( )  

    Objective To investigate the neurofilament protein (NF) and synaptophysin(SYN) distribution in the developing stage of human embryonic spinal cord and their clinical significance. Methods Totally 16 of human embryos, aged from 2 to 4 months were used in this study. Immunohistochemical staining was uesd to detect the expression and distribution of NF and SYN in the spinal cord. Results At the second month stage, NF expression was weakly positive in only the marginal layer on both sides of the spinal cord. At 3-4 months, NF expression was positive or strong positive in the spinal cord center crack before and after the middle of partition and cortex. In the ependymal layer of the lateral part of the expression of NF positive nerve fibers were seen in the gray matter. From 2 to 4 month-age, all visible SYN positive expressions distributed in the spinal cord. With the increase of gestational age, SYN positive expression in the spinal cord staining intensity value was increased first, and then decreased. The number of positive values increased month by month. Conclusion NF and SYN both take part in the regulation of human embryonic spinal cord tissue growth.

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    Expression of the receptor for advanced glycation end products in mice with chronic adriamycin-induced cardiotoxicity
    SHI Sheng-feng ZHU Jin-zhou ZHANG Rui-yan*
    2014, 45 (4):  550-554.  doi: 10.3969/j.issn.0529-1356.2014.04.020
    Abstract ( )  

    Objective To study the expression of receptor for advanced glycation end products (RAGE) in chronic adriamycin (ADR)-induced cardiotoxicity mice. Method A mouse model of chronic cardiotoxicity was established by intraperitoneal injection of 5 mg/kg ADR once a week for continuous 3 weeks. Mice treated with the same volume of saline were used as control group (6 mice in each group). Echocardiography and histopathological examination were used to evaluate the myocardial injury, cardiac function and ventricular remodeling. The expression of RAGE in cardiac tissues was detected by immunohistochemical staining and Western blotting. Results Echocardiography indicated that ADR-treated mice had reduced left ventricular ejection fraction, while histopathological analysis of cardiac tissues showed obvious cardiac damage. RAGE protein in cardiac tissues was highly expressed in chronic ADR-induced cardiotoxicity mice. Conclusion RAGE is highly induced and may be involved in the cardiotoxicity after ADR treatment.

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    Different distribution and expression of mammalian target of rapamycin complex in the kidney of diabetic nephropathy mice
    ZHAO Hong JI Qian-qian LI Yong-xia DUAN Qiu-hong YAO Li-jun*
    2014, 45 (4):  555-560.  doi: 10.3969/j.issn.0529-1356.2014.04.021
    Abstract ( )  

    Objective To investigate the different distribution and expression of mammalian target of rapamycin complex (mTORC) in the kidney of diabetic nephropathy (DN) mice. Methods Fourteen eight-week-old male C57BL/6 mice were assigned to 2 groups: the control group (n=7) and the streptozotocin (STZ)-induced DN group (n=7). Blood and urinary variables including glucose, albumin, creatinine and albumin/creatinine ratio were assessed 2 weeks after STZ injection. Hematoxylin-eosin staining was performed for renal pathological analyses. The distributions of mTOR, phosph-ser2448-mTOR(p-mTOR), mTORC1(Raptor), mTORC2(Rictor) and phosph-ser240/244-S6K1 (p-S6K1) were determined by immunofluorescence. The expression levels of mTOR, p-mTOR, mTORC1(Raptor), mTORC2(Rictor), S6K1 and p-S6K1 were detected by Western blotting. Results Two weeks after STZ injection, the diabetic mice developed albuminuria (P<0.01) and renal hypertrophy (P<0.05). The immunofluorescence positive staining for mTOR, Raptor, and Rictor was distributed in the epithelial cells of proximal tubules, glomerular mesangium and capillary loops as well as the medullary collecting ducts of the control mouse kidney. These positive signals increased in the DN mouse kidney (P<0.05). However, pS6K1 was not detected in the inner medulla of control mouse and p-mTOR was not found in the glomeruli of both control and DN mice. Conclusion mTORC is widely expessed in the mouse kidney and participates in the development of DN, whereas the 2448 serine phosphorylation of mTOR may be not implicated in the hyperglycemia mediated glomerular injury.

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    Biocompatibility of rat’s nature decellularized pancreatic biological scaffolds
    SHAO Ying-kuan YAN Xia-lin RAO Zhi-heng HUANG Gao-jian LI Jia-wei HUANG Jun-jie MEI Jin* LIN Ke-zhi*
    2014, 45 (4):  561-568.  doi: 10.3969/j.issn.0529-1356.2014.04.022
    Abstract ( )  

     Objective To harvest pancreatic tissues from rats, prepare decellularized bio-derived pancreatic scaffolds (DBPS), and to examine the integrity and biocompatibility of the scaffolds. Methods Normal pancreases were harvested from healthy adult SD rats. DBPS was prepared by perfusing SDS and Triton X-100 through bile duct and the portal vein, respectively. After decellularization, normal pancreatic tissue and DBPS were compared via HE staining, and transmission electron microscopy (TEM). Abdominal wall and subcutaneous implantations were used to compare biocompatibility, and the remain quantity of residual protein and growth factors were determined via enzyme linked immunosorbent assay(ELISA). MTT assay was used to test the scaffolds’ cytotoxicity. The scaffolds were co-cultured with endotheliocyte. Results HE staining and TEM study indicated no residual cells in the DBPS as well as preservation of the complete extracellular matrix. The remain quantity of residual protein and growth factors in ECM was high. The abdominal wall and subcutaneous implantation revealed that DBPS triggered a lower immune response as compared to the control group. MTT assay showed little cytotoxicity. Endotheliocyte assembled and growed with the scaffolds together.
    Conclusion DBPS are completely decellularized, and exhibit a higher level of biocompatibility in vivo. Using the way of vessels can make the integrity of extracellular matrix to be fully preserves and contain more growth factors. So using vessels way is better than bile duct.

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    In situhybridization and analysis of core sequence of follicle-stimulating hormone and its receptor in rats submaxillary
    YU Hui*LIU Gui-yun 
    2014, 45 (4):  569-572.  doi: 10.3969/j.issn.0529-1356.2014.04.023
    Abstract ( )  

    Objective To explore if the expression of follicle-stimulating hormone (FSH) and its receptor (FSHR) exist in the rat submandibular gland tissue, which may provide a theoretic evidence for further studying on adjusting function of the FSH in submaxillary gland. Methods Twenty normal SD rats, with a mean body weight of (200±20)g, were adopted to collect out the submandibular gland after abdominal cavity anesthesia. The method ofin situ hybridization was employed on cellular localization of FSH and FSHR. RT-PCR was used to detect if FSH and its receptor mRNA exist in the rat submandibular gland. RNA was extracted from the submandibular gland, then FSH and the core cDNA of its receptor gene were obtained with reverse transcription polymerase chain reaction and analyzed on its sequence. Results The epithelial cells of serous acinus and granular convoluted tubule in submaxillary gland of rats had FSH and FSHR mRNA hybridized signals, which were detected in cytoplasm with negative nuclei. Specific bands of FSH cDNA and FSHR cDNA were identical to be 193bp and 413bp respectively. Conclusion FSH and FSHR can be synthesized by the epithelial cells of serous acinus and granular convoluted tubule in submaxillary gland of rats, thus this study further proves that the submandibular gland is the target organ of FSH.

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    Genetic relationship among twenty-six populations using Y-STR polymorphism information
    YANG Xin SUN Hong-bing* LUO Ji-huai HA Fei ZHANG Zi-long
    2014, 45 (4):  573-577.  doi: 10.3969/j.issn.0529-1356.2014.04.024
    Abstract ( )  

    Objective To study the genetic relationship of the Y chromosomal short tandem repeat gene loci in Lanzhou Han population and other 25 populations. Methods The frequency of alleles of Y-STRloci was obtained from a sample of 500 unrelated individuals living in Lanzhou City, and other 25 populations in different areas collected from the published data were used to calculate the genetic similarity coefficient and genetic distance. Phylogenetic trees based on the genetic distance were established. Results Populations of Lanzhou, Beijing, Shanxi and Inner Mongolia were in an identical cluster. Compared with minorities, the genetic distance between Lanzhou Han population and Inner Mongolia Mongolian population was dramatically smaller from other subpopulations. The populations in Malays and Indians were far from the other groups. Conclusion The Y-STR gene frequency distribution in 26 populations has identified differentiation in race, clime and evolution, and it is basically identical with the classification of human races which is similar to or according with other molecular anthropology research conclusions.

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    Skinfold thickness of Han adults in Jiangsu province
    ZHANG Xing-hua ZHENG Lian-bin* YU Ke-li ZHAO Da-peng WANG Zhi-bo WANG Yang RONG Wen-guo ZHANG Xiao-rui
    2014, 45 (4):  578-581.  doi: 10.3969/j.issn.0529-1356.2014.04.025
    Abstract ( )  

    Objective To study the characteristics of skinfold thickness of Han adults in Jiangsu province. Methods The skinfold thicknesses of facial, subscapular, suprailiac, biceps, triceps and calf on 311 urban adults (157 males and 154 females) and 421 rural adults (213 males and 208 females)of Han were investigated in Huaian city of Jiangsu province. Results The thickness of skinfold of urban females were thicker than that of urban males. Rural adults were the same. Han adults of Jiangsu showed the most significant differences between urban areas and rural areas. The values of six skinfold thicknesses of Jiangsu urban adults have positive correlation with age. Conclusion Han adults of Jiangsu show the most significant differences between genders.

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    Application of Cell Counting Kit-8 in detecting the growth inhibiting effect of 5-Aza-2’-deoxycytidine on chronic myeloid leukemia cells
    LIU Yue SHUAI Chun LI Jie-sheng YIN Hong SONG Yan-bin MA Wen-li*
    2014, 45 (4):  582-584.  doi: 10.3969/j.issn.0529-1356.2014.04.026
    Abstract ( )  

      Objective To study the application of Cell Counting Kit-8(CCK-8) in detecting the growth inhibiting effect of 5-Aza-2’-deoxycytidine on chronic myeloid leukemia cell.
    Methods The proliferation of K562 cells was detected by CCK-8 with different concentrations of 5-Aza-2’-deoxycytidine and the cell cycle and apoptosis of K562 cells were detected after K562 treated by 50% inhibitory concentration of 5-Aza-2’-deoxycytidine. Results The 50% inhibitory concentration of 5-Aza-2’-deoxycytidine was 15.55nmol/L, after treated with this concentration, K562 cells showed that G2 phase arrest occurred, proliferation inhibited and apoptosis peaks appeared. Conclusion Inhibition of proliferation of K562 cells with different concentrations of 5-Aza-2’-deoxycytidine varied in a dose-dependent relationship, and 5-Aza-2’-deoxycytidine could promote apoptosis of K562 cells. CCK-8 can be used in detecting the effect of 5-Aza-2’-deoxycytidine on chronic myeloid leukemia cells.

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