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2014, Volume 45 Issue 5
06 October 2014 Previous Issue    Next Issue

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Effects of Jagged1 on hippocampal radial glial cells’proliferation and neuronal differentiation QIN Jian-bing CHENG Min JIN Guo-hua* LI Hao-ming SHI Jin-hong ZOU Lin-qing TIAN Mei-ling 2014, 45 (5):  585-590.  doi: 10.3969/j.issn.0529-1356.2014.05.001 Abstract ( )   Objective To investigate the effect of Jagged1 on hippocampal radial glial cells (RGCs) proliferation and neuronal differentiation in vitro.Methods Hippocampal RGCs were cultured in vitro, the agonist Jagged1 and(or) inhibitor DAPT of Notch signaling were added into the culture medium, and then the cells were divided into control group, Jagged1 group, Jagged1 combined with DAPT group and DAPT group. CCK-8 regent was used to detect cells’ vitality; immunofluorescent was used to detect the number of BLBP/Ki67 double labeled cells and differentiated microtubule associated protein-2(MAP-2) positive cells. Results Cell vitality in Jagged1 group was obviously higher than that of the other groups. The number of BLBP/Ki67 double labeled cells and differentiated MAP-2 positive cells were more than other groups. Conclusion Jagged1 promotes the proliferation and neuronal differentiation of hippocampal RGCs in vitro. References | Related Articles | Metrics

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Expression of natriuretic peptide receptor A in the developing retina of the mouse LI Jin-ju LI Rui-ling LI Xue LIU Kai Deng Jie-xin WU Ping* DENG Jin-bo* 2014, 45 (5):  591-598.  doi: 10.3969/j.issn.0529-1356.2014.05.002 Abstract ( )   Objective Our purpose is to investigate the expression of natriuretic peptide receptor A (NPR-A) in the retina and to understand the NPR-A’s functions during the mouse development. Methods Mice eyes were harvested from E16 (embryonic day 16) to P90 (postnatal day 90). Total of 127 eyes were used in the study. Immunohistochemistries of NPR-A were carried out. Results During development, NPR-A was widely expressed in the retinal neurons. In the outer nuclear layer, NPR-A began to appear in the inner and outer projections of cone and rod cells at P7, but decreased at P14. From P30 afterward, it continued to express weakly. In the inner nuclear layer, NPR-A expressed in the dendrites of bipolar cells weakly from P7 to adulthood, whereas no expression in horizontal cells. In the ganglion cell layer, NPR-A started highly to express in the ganglion cell bodies at E16, and in the meantime, in the nerve fiber layer, ganglion cell axons, NPR-A was expressed highly from embryonic to adult. In the inner and outer plexiform layers, NPR-A was highly expressed at P14, but decreased gradually after P30. In addition, NPR-A also widely expressed in the inner protrusions of Müller cells. Conclusion NPR-A participates in the development of the retina, and may be the key molecule in the developing retinal neurons. Moreover, it plays an important regulatory role in the functional activity of Müller cells. References | Related Articles | Metrics

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Long-term evaluation of functional recovery and nerve regeneration following tubulation repair of nerve defects in mice MI Da-guo ZHANG Yan-ping GU Tian-wen ZHAO Ya-hong HU Wen* 2014, 45 (5):  599-604.  doi: 10.3969/j.issn.0529-1356.2014.05.003 Abstract ( )   Objective This study is to identify long-term functional recovery and maturity of regenerated nerve fibers after repairing mouse nerve defects with chitosan/polylactide-co-polyglycolide artificial nerve grafts (CPANGs). Methods Mouse sciatic nerve defects, 2mm in length, were bridged by CPANGs (n=6), with nerve autograft (n=6) and nerve defect (n=6) as controls. Plantar test, electrophysiological examination and laser Doppler perfusion imaging following nerve crush were carried out at 1 year after repair to assess nerve function recovery, while muscle wet weight ratio, histological assessment and transmission electron microscopy were performed to evaluate nerve re-innervation and maturity of regenerated nerve fibers. Results When compared to the autograft group, the CPANG group did not show statistically significant difference in functional recovery in terms of paw withdrawal latency, neurogenic vasodilatation, amplitude and latency of compound muscle action potentials (CMAPs), wet weight ratio of gastrocnemius and tibialis cranialis muscles, number of myelinated nerve fibers and density of unmyelinated axons. However, both these two repair groups exhibited significantly longer CMAPs latency, thinner myelin sheath and a lagbehind shift of diameter distribution of myelinated axons as compared to the normal control. Conclusion At 1 year after the mouse sciatic nerve defect was repaired by CPANGs, sensory and autonomic nerve function, number of regenerated axons and muscle re-nnervation degree were recovered to the same extent as nerve autografting, but the regenerated nerve fibers were in a state of immaturity. References | Related Articles | Metrics

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Comparison of glucose and lipid metabolism in two types of Parkinson’s disease rat models MENG Xiang-zhi ZHENG Rui-pan ZHANG Ying QIAO Ming-liang JING Peng GAO Yan* 2014, 45 (5):  605-609.  doi: 10.3969/j.issn.0529-1356.2014.05.004 Abstract ( )   Objective To select an ideal Parkinson’s disease (PD) animal model with metabolic abnormalities for subsequent experimental studies. Methods A total of 62 Sprague-Dawley male rats were randomly divided into four groups: damaged medial forebrain bundle (MFB) model group, damaged medial forebrain bundle (MFB) sham group, damaged Striatum model group and damaged Striatum sham group. After detecting the rotation experiment, successful model rats of two groups were selected to detect the changes of food intake, body weight, blood glucose and intra-abdominal adipose tissue. Results It was easier to produce a PD model by destroying MFB than striatum. Compared with sham-operated rats, MFB model rats showed significant abnormality both in reduction of body weight ［(218.1±13.99)g vs (252.7±10.1)g, P<0.05］ and high blood glucose appeared at 15min and 30min after introperitoneal glucose tolerance test (IPGTT). Their perirenal white adipose tissue was significantly reduced (both left and right side). Striatum model rats only appeared decreased food intake ［(13.95±0.25)g vs (20.23±0.86)g, P<0.001］ and impaired glucose regulation at 15min, 30min and 60min after IPGTT. Their body weight and adipose tissue did not change significantly. Conclusion No matter in the success rate or metabolismrelated indicators, MFB damaged rat model of PD is more suitable to study PD patients with abnormal lipid metabolism compared with Striatum rat model. References | Related Articles | Metrics

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Comparisons of expression of phosphorylated extracellular signal-regulated kinase in different levels of anterior cingulate cortex in the rat with neuropathic pain FANG Fang SHAO Xiao-mei SHEN Zui SUN Jing FANG Jian-qiao* 2014, 45 (5):  610-615.  doi: 10.3969/j.issn.0529-1356.2014.05.005 Abstract ( )   Objective To observe the distribution features of phosphorylated p44/42 extracellular signal-regulated kinase(p-ERK) induced by affective pain in the anterior cingulate cortex (ACC) of the rat. Methods Twelve male SD rats were randomly divided into the control group (n=6) and spinal nerve ligation (SNL) group (n=6). The rats of the SNL group were exposed to unilateral L5 spinal nerve ligation (SNL) surgery. The behavior was examined by mechanical pain thresholds (MPTs) and the pain affect was measured using open-field test, elevated zero maze. The expressions of p-ERK in ACC (AP 3.2,2.7,2.2mm from Bregma) were detected with immunofluorescence test. Results In comparison with the control group, the unilateral mechanical pain threshold in the SNL group decreased significantly after operation. The operation also resulted in pronounced anxiety-like behavior. The p-ERK labelled cells in different levels of ACC(AP 3.2,2.7,2.2mm from Bregma) of SNL rats were (11.89±2.57),(32±4.67 ) and (17.56±2.04 ), respectively, while the control group rats were (12.44±2.16),(10±0.87 ) and (10.11±1.36). These data indicated that compared with the control group, the expressions of p-ERK in ACC(AP 2.7,2.2mm from Bregma) increased significantly in the SNL group after operation(P<0.01), besides for the level of ACC(AP 3.2mm from Bregma) (P>0.05). Conclusion The results suggest that the neuropathic pain can result in anxiety-like behavior and increased expressions of p-ERK in ACC of the rat. This change may be closely related to the expressions of p-ERK in ACC(AP 2.7，2.2mm from Bregma), but not to the level of ACC(AP 3.2mm from Bregma). References | Related Articles | Metrics

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Ativation of gliacytes and p38 mitogen-activated protein kinase and possible mechanism of neuronal apoptosis induced by Aβ25-35 injection into hippocampus in rats WANG Yuan-wei ZHENG Guan-yi* CHEN Xiao-chun ZHANG Jing HUANG Tian-wen YE Hong PAN Xiao-dong 2014, 45 (5):  616-621.  doi: 10.3969/j.issn.0529-1356.2014.05.006 Abstract ( )   Objective To investigate the relationship between activation of gliacytes, mitogen-activated protein kinase (p38MAPK) and neuronal apoptosis after microinjecting aggregated Aβ25-35 into hippocampus. Methods The model was established by using stereotaxic technique to inject 10μg aggregated Aβ25-35 into dorsal hippocampus in rats. The rats were grouped as the control, vehicle and model groups. Immunohistochemistry and Western blotting were used for detection of activation of microglia(MG), atrocytes (AS) and expression of p-p38MAPK in the hippocampus. ELISA was used to evaluate the level of TNF-α and IL-1β. The survival neurons were observed by Nissl staining and the apoptotic neurons were identified by tunnel staining. Results Expression of ox-42, GFAP, p-p38MAPK were up-regulated in hippocampus, as well as TNF-α、IL-1β, which reached a highest value on the 7th day after injection of Aβ25-35. However, the number of neuron with Nissl positive decreased gradually, and the tunnel positive neurons increased highly and reached a peak value on the 7th day.There were significant differences between the control and vehicle group（P<0.01）. Conclusion Apoptosis of the neuron caused by Aβ25-35 injection may result from activation of gliacytes, p38 MAPK and increase of TNF-α and IL-1β level. References | Related Articles | Metrics

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Effect of 3-N-butylphthalide pretreatment on the score of neurological deficit, oxidative stress and pathomorphology in rats with cerebral ischemia reperfusion injury JI Hai-ru KONG Ling-wei KONG Wei ZHAO Shu-min* ZHENG Xiao-ying CHEN Meng 2014, 45 (5):  622-626.  doi: 10.3969/j.issn.0529-1356.2014.05.007 Abstract ( )   Objective To investigate the effects of 3-N-butylphthalide (NBP) pretreatment on the score of neurological deficit, oxidative stress and pathomorphology in rats with cerebral ischemia reperfusion injury (CIRI). Methods Ninety male SD rats were randomly divided into sham operation group (Sham group), model group (IR group), NBP pretreatment low dose group (NBPⅠgroup), NBP pretreatment middle dose group(NBPⅡgroup) and NBP pretreatment high dose group(NBPⅢ group), 18 rats per group. Pretreatment was given once a day within 1 week before establishing the model of cerebral ischemia reperfusion injury. The model of middle cerebral artery occlusion(MCAO) was subjected by suture method. The score of neurological deficit was executed after ischemia for 2h and reperfusion for 24h in all the rats. The cerebral infarction was observed by TTC staining. The pathologic change of brain was observed by HE staining under the microscope. Hydroxylamine method was used to detect activity of SOD, chemical colorimetry method was used to measure activity of GSH-PX, and TBA method was used to detect content of MDA. Results (1) In Sham group, the score of neurological deficit and the percentage of infarction volume were zero, the morphology of nerve cell was regular, and activity of SOD, GSH-PX and content of MDA of brain tissue were normal. (2) Compared with IR group, the score of neurological deficit was significantly reduced in NBP pretreatment groups (all P＜0.01); the score of neurological deficit was decreased progressively in turn in NBP Ⅰ,Ⅱ,Ⅲ group (all P＜0.05). (3) Compared with IR group, the percentage of infarction volume was cut down progressively in turn in NBPⅠ,Ⅱ,Ⅲ group (all P＜0.05), and neuron injury was also induced obviously in NBP pretreatment groups.(4) Activity of SOD, GSH-PX was largely increased, and content of MDA was greatly decreased in NBP pretreatment groups(P＜0.01). Activity of SOD, GSH-PX went up progressively in turn, and contents of MDA were cut down progressively in turn in NBP Ⅰ,Ⅱ,Ⅲgroup (all P＜0.05). Conclusion 3-N-butylphthalide can significantly up-regulate the activity of SOD and GSH-PX, decrease the content of MDA, reduce the percentage of infarction volume, and relieve the damage of nerve cell to preventively protect the rats with cerebral ischemia reperfusion injury. References | Related Articles | Metrics

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Effect of estradiol on proliferation of rat hippocampal neural stem cells LIU Xiao-dong ZHANG Xia-nan HAO Ning JU Qian-qian QIN Jian-bing TIAN Mei-ling JIN Guo-hua Lü Guang-ming* 2014, 45 (5):  627-632.  doi: 10.3969/j.issn.0529-1356.2014.05.008 Abstract ( )   Objective The aim is to observe the role and mechanism of estradiol (E2) on the proliferation of rat hippocampal neural stem cells (NSCs). Methods Twenty hippocampi from embryonic 17-day (E17) SD rats were dissociated and plated into culture flasks with NSCs specific medium containing different concentrations of estradiol. The proliferation and the vitality of NSCs were detected by immunofluorescence against BrdU and MTT assay. The expression of estrogen receptors (ERα and ERβ) was measured by immunofluorescence staining combined with Nestin double labeling. Results BrdU and MTT assay results showed that the cell number increased when the concentration of estradiol increased from 10-10 to 10-8 mol/L. The number of cell proliferation and the viability of cells were best at the concentration of 10-8 mol/L compared to the other groups. However, when the estradiol concentration was increased from 10-8 to 10-6 mol/L, the cell proliferative capacity declined gradually. Double immunofluorescence labeling showed that the two types of estrogen receptors (ERα and ERβ) were expressed in the cultured hippocampal NSCs. Conclusion Estradiol promotes the proliferation of hippocampal NSCs in a certain concentration range, and ERα and ERβ may be involved in the estradiol-induced proliferation. References | Related Articles | Metrics

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Regulating effect of berberine on macrophage phenotype transformation in hepatic tissue of mice with methionine-choline deficiency diet induced non-alcoholic steatohepatitis ZHANG Hui-qin LIU Ze-zhou XU Chang LIU Xin LOU Jin-li LI Jian NIU Jian-zhao HAO Yu * 2014, 45 (5):  633-638.  doi: 10.3969/j.issn.0529-1356.2014.05.009 Abstract ( )   Objective To determine the efficacy of berberine in the treatment of non-alcoholic steatohepatitis (NASH), and to investigate the regulating effect on macrophage phenotype transformation in hepatic tissue on methionine-choline deficiency (MCD) diet induced NASH mice. Methods Fourty male C57BL/6 mice were randomly divided into 4 groups (10 mice per group): the normal group (fed with normal diet), the NASH model group (fed with MCD diet), rosiglitazone treatment group (30mg/kg) and berberine treatment group (150mg/kg). Drugs were adopted in the preventive intervention method for 2 weeks. The hepatic histopathological method was adopted to evaluate the drug therapeutic effect. The serum levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-6, and IL-10 were examined with ELISA method. M1 and M2 phenotype were detected by flow cytometry. Results The results showed berberine improved the degree of hepatic histopathology. Berberine not only reduced the level of TNF-α, but also increased the level of IL-10 in serum on NASH mice significantly (P<0.05). Flow cytometry data indicated that berberine decreased M1 type macrophages and increased M2 type macrophages in liver tissue. The ratio of M1/M2 was significantly decreased in berberine and rosiglitazone treated group (P<0.01). Conclusion Berberine may improve the hepatic pathological process in MCD diet induced NASH model possibly through modulating macrophage phenotype transformation, i.e. The ratio of M2 type is more than M1 type in hepatic tissue, and increasing anti-inflammatory cytokines. References | Related Articles | Metrics

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Effect of stromal cell derived factor-1α/CXCR4/CXCR7 axis on migration of the bone marrow mesenchymal stem cells WANG Yu-lan HE Xiao-mei* TANG Wei GU Yi-ping ZHANG Shi-chun TANG Man XU Tie-jun GAO Dian-shuai 2014, 45 (5):  639-645.  doi: 10.3969/j.issn.0529-1356.2014.05.010 Abstract ( )   Objective To investigate expression of CXCR4 and CXCR7 protein and mRNA, which are the receptors of stromal cell derived factor-1α(SDF-1α), in the bone marrow mesenchymal stem cells(BMSCs); to explore the role of SDF-1α/CXCR4/CXCR7 axis in migration of BMSCs in vitro and the possible mechanism. Method BMSCs were isolated from rats and cultured in vitro. CD29, CD44 and CD34 of the cells were identified by flow cytometry. CXCR4-selective antagonist AMD3100 and CXCR7-specific neutralizing antibody were applied to block CXCR4 and CXCR7 respectively. The expressions of CXCR4 and CXCR7 mRNA and protein on BMSCs were detected with RT-PCR and Western blotting. Transwells chamber test was used to observe the migration of BMSCs. The BMSCs were divided into the BMSCs group(A), the AMD3100 pretreated BMSCs group(B), the CXCR7-specific neutralizing antibody pretreated BMSCs group(C), the AMD3100+CXCR7-specific neutralizing antibody pretreated BMSCs group(D). Result Flow cytometry showed that the expressions of CD44 and CD29 were positive, while the expression of CD34 was negative in the third passage of BMSCs(P3-BMSCs). CXCR4 and CXCR7 protein and mRNA were both expressed in P3-BMSCs. Compared with the A group, the expression of CXCR4 and CXCR7 protein declined significantly in the B group and the D group; the protein expression of CXCR7 in the C group was lower compared with the A group(P<0.05). However, the expression of CXCR4 mRNA and CXCR7 mRNA had no significant difference between groups. SDF-1α factor promoted migration of BMSCs(P<0.05). Compared with the 0μg/L group, the numbers of migrated cells were increased significantly in both of the 10μg/L group and the 100μg/L group(P<0.01). The number of migration of BMSCs was significantly higher in the 100μg/L group than that of the 10μg/L group(P<0.01). AMD3100 and CXCR7-specific neutralizing antibody both inhibited significantly the migration of BMSCs(P<0.05), and the attenuate effect was more significant when they worked together(P<0.05). Conclusion CXCR4 and CXCR7 receptors are coexpressed in P3-BMSCs; the SDF-1α factor can promote the migration of BMSCs in the concentration dependent manner; SDF-1α/CXCR4/CXCR7 axis is involved in the migration of BMSCs, and both of the CXCR4 and CXCR7 receptors have a synergistic promoting effect to the BMSCs migration. References | Related Articles | Metrics

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Expression of microtubule-associated protein 1 light chain 3 and Beclin-1 in alveolar macrophages of silicosis rats ZHANG Juan ZHAO Man-man LI Ran ZHU Hui-xing TIAN Yan-xia CUI Jian-zhong GAO Jun-ling* 2014, 45 (5):  646-651.  doi: 10.3969/j.issn.0529-1356.2014.05.011 Abstract ( )   Objective To observe expression of autophagy-related proteins LC-3 and Beclin-1 in alveolar macrophages in the silicosis model of rats, and to investigate the molecular mechanisms of silicosis formation from cells autophagy perspective. Methods Fifty adult male SD rats were randomly divided into control and model groups, 25 rats for each group. The silicosis model was made by one-time infusion of silica dust suspension through the trachea without exposed. The rats were sacrificed on the 1st, 3rd, 7th, 14th or 28th day after the modeling. Alveolar macrophages were obtained from bronchoalveolar lavage and used for subsequent research after culture and purification. Morphological characteristics of alveolar macrophages were observed by HE staining and transmission electron microscope; The expression of LC-3 and Beclin-1 was detected by means of immunocytochemistry and Western blotting in each group.  Results Compared with the control group, alveolar macrophages of the model group had larger volume and abundant cytoplasm, the phagocytic silica dust particles were observed in some cells, and autophagosomes were detected by transmission electron microscope. The expressions of LC-3 and Beclin-1 were increased at all time points in the model group(P<0.05). Both LC-3 and Beclin-1 began to increase at the 1st day. As the extension of time the expression gradually enhanced, peaked at the 14th day(P<0.05), and decreased at the 28th day, but higher than the basal expression. Conclusion Autophagy is activated in alveolar macrophages of the silicosis model, and alveolar macrophages autophagy is involved in the pathological process of silicosis in the rat.   References | Related Articles | Metrics

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Expression changes of islet amyloid polypeptide, somatostatin positive cells of pancreatic islet in type 1 diabetes mellitus mice LI Rong-rong LIANG Wen-mei* 2014, 45 (5):  652-655.  doi: 10.3969/j.issn.0529-1356.2014.05.012 Abstract ( )   Objective To explore the changes of the expression of islet amylodi polypeptide (IAPP) and somatostatin(SS) of islet in type 1 diabetes mice, and the mechanism of the expression changes. Methods We established the diabetes model in C57BL/6J mice by low-dose streptozotocin (STZ) injection. The excisions of the pancreas tails removed on the 3th, 7th, 10th, 14th, 21th and 28th day. Tissues were assessed by immunohistochemical SABC, immunofluorescence in the study. Results 1. Numerical density on area (NA) of IAPP positive cells in experimental group (EG) decreased since the 3th day, but average absorbance increased since the 7th day. 2. NA of SS-IR cells in EG increased since the 3th day, and average absorbance increased since the 7th day. 3.The results of immunofluorescence double staining showed that, IAPP and SS could coexpression in part of cells in islets. Conclusion The number of IAPP positive cells in type 1 diabetes is decreased, but the immunoreaction increased. Immunoreaction and number of SS positive cells increase. Both of them are involved in the pathogenesis of type 1 diabetes. References | Related Articles | Metrics

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Effect of lipopolysaccharide on the iron metabolism in macrophages WANG Li JIANG Bao-hua QIAN Zhong-ming DUAN Xiang-lin* 2014, 45 (5):  656-662.  doi: 10.3969/j.issn.0529-1356.2014.05.013 Abstract ( )   Objective To study the effect of lipopolysaccharide(LPS) on the activity of primary cultured macrophages and the distribution of divalent metal transporter 1 (DMT1) and ferroportin 1 (FPN1). Methods Primary cell culture, MTT chromotest, cytochemistry chromotest and cell immunofluorescence techniques were used in this work. Results DMT1 was mainly distributed in the cytoplasm, which illuminates that DMT1 mediates the macrophage intracellular transit of iron from phagolysosome to cytoplasm. FPN1 was distributed in the cytoplasm and membrane, and the cytoplasm was the main site of FPN1 distribution in macrophages. Conclusion Iron liberation from heme inside the phagolysosome occurs after erythrophagocytosis and it is possible that FPN1 mediates intracellular transit of iron released by heme catabolism. The study found that LPS promoted the cell growth and this effect was reached to the peak in the 10-5 μg/L LPS group, but the cell growth was blocked with the increase of LPS concentration. References | Related Articles | Metrics

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Induction effect of hepatocyte growth factor and insulin-like growth factor on cardiac stem cells ZHU Bei-bei CAI Xin-hua SUN Yin-ping* 2014, 45 (5):  663-669.  doi: 10.3969/j.issn.0529-1356.2014.05.014 Abstract ( )   Objective To investigate whether hepatocyte growth factor (HGF) and insulin-like growth factor (IGF1) induce cardiac stem cells (CSCs) to proliferate and directly differentiate into cardiomyocytesin vitro. Methods The myocardial tissues were dissected for primary culture of CSCs with the method of explants. The expressions of c-kit and CD34 were examined with immunofluorescence. Primary cells were purified with c-kit by flow cytometry. CFDA SE fluorescent probe was used to detect the proliferation of c-kit+CSCs. C-kit+CSCs were divided into two groups, and cardiac stem cells group and co-cultured with cardiomyocytes group, both group were cultured with HGF and IGF1. An inverted microscope was used to observe changes in cell number and morphology in different periods. Living cells workstation was used to observe CFDA SE fluorescence intensity, to acquire images and do statistical analysis. Immunofluorescence technique was used to detect the expression of Nkx2.5 and cardiac troponin T. Results In cardiac stem cells group，CSCs had no obvious changes in cell number. In co-cultured with cardiomyocytes group, CSCs proliferated and had changes in morphology. Nkx2.5 and cTnT were positively expressed. Several CSCs differentiated into beating cardiomyocytes. Conclusion In co-cultured with cardiomyocytes condition, HGF and IGF1 may promote CSCs to proliferate and differentiate into beating cardiomyocytes. References | Related Articles | Metrics

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Effects of p38-2G4 high-expression on the proliferation and erythriod differentiateon of murine erythroleukemia cells ZHU Xiao-fang SHI Ming-ming YANG Zu-li ZHAO Fu-kun ZHANG Shi-fu* 2014, 45 (5):  670-674.  doi: 10.3969/j.issn.0529-1356.2014.05.015 Abstract ( )   Objective To explore the effect of mouse proliferation-associated protein 2G4 (p38-2G4) high-expression on the proliferation and erythriod differentiation of murine erythroleukemia (MEL) cells. Methods To establish the recombinant lentivirus vector p38-2G4-pLJM1, the p38-2G4-pLJM1 was cotransfected into HEK293T cells to obtain lentivirus with pCMV-VSV-G and pCMV-dR8.2. Lentivirus were infected into MEL cells to establish the stably p38-2G4 high-expressed MEL cells. Western blotting was used to analyse the high-expression efficiency. MTT assay and benzidine staining were applied to detect the cell viability and hemoglobin synthesis of the stable cell line in presence/absence of inducers. Results Western blotting showed that the p38-2G4 high-expression stable cell strain had a higher expression of p38-2G4 as compared to the control group (MEL)（P<0.05）. MTT result showed that there was no difference between the p38-2G4 high-expression cell strain and the control group（P>0.05）, but the hemoglobin synthesis had been reduced as compared to the control group（P<0.05）. Conclusion p38-2G4 high-expression does not affect the cell viability of MEL cells, but inhibits the erythriod differentiation of MEL cells in three independent experiments. References | Related Articles | Metrics

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Expression of fibroblast growth factor-2 and fibroblast growth factor receptor-4 in human papillary thyroid carcinoma and their significances LI Qian LI Li GUO Shu-qin* ZHANG Yun-liang KONG Fan-qiang LI Fei KANG Jun-peng WU Jing-fang GAO Fu-lu 2014, 45 (5):  675-681.  doi: 10.3969/j.issn.0529-1356.2014.05.016 Abstract ( )   Objective To investigate the expression of fibroblast growth factor-2 (FGF-2) and fibroblast growth factor receptor-4(FGFR-4) in the papillary thyroid carcinomas（PTC）and clinical significance. Methods Immunohistochemistry and Western blotting for the expression of FGF-2 and FGFR-4 were performed in 89 cases of PTC and 30 cases of normal thyroid tissues (NTT) adjacent to the tumors. Results Immunohistochemistry results showed that, FGF-2 and FGFR-4 expressions were high in thyroid carcinoma (P<0.01，P<0.01)in contrast to that in the normal thyroid tissues, and the difference was statistically significant;There was a positive linear correlation between expressions of FGF-2 and FGFR-4 and lymph node metastasis (χ2=14.798,P<0.01;χ2=7.27,P<0.01)and differentiation degree (χ2=13.824,P<0.01;χ2=16.921, P<0.01) in papillary thyroid carcinoma,while there was no difference in gender,age and tumor size(P>0.05). Analyzed by Western blotting technique,FGF-2 and FGFR-4 expressions in thyroid carcinoma were significantly higher than that in normal tissue,with decrease of cancer degree of tissue differentiation and significantly up regulated expression (P< 0.05). Expressions of FGF-2 and FGFR-4 were in a positive linear correlation in the disease（rs=0.434,P<0.01）. Conclusion The expressions of FGF-2 and FGFR-4 are correlated with papillary thyroid cancer and they participated in the process of invasion and metastasis, both of which have a positive synergistic effect. The degree of malignancy and biological behavior are meaningful and comprehensive indicators,which provide a theoretical basis for the subsequent experimental studies of cellular and molecular biology. References | Related Articles | Metrics

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Anatomical symmetry of the intrahepatic Glisson system YU Zhi-yong* LIU Qiao-yun WEI Jia MA Xin-yi GAO Ze-hai CAI Qiang 2014, 45 (5):  682-687.  doi: 10.3969/j.issn.0529-1356.2014.05.017 Abstract ( )   Objective To observe the anatomical symmetry of the structures and istribution of the Glissonean pedicle of the intrahepatic Glisson system，integrating with embryology and comparative anatomy. Methods Morphology of the Glissonean pedicle of liver was examined through peeling and dissecting 20 adult corpses without liver pathological changes. The relevant data were collected and analyzed statistically. Meanwhile, we tried to elucidate elaborating the symmetry theory of liver anatomy through the dissection anatomy,embryonic anatomy and comparative anatomy. Results The angle between main stem of Glisson system/left branch of Glisson system(GM/GL) was (76.7±17.36)°. The angle between GM/GR was (81.4±13.8)°. The length of the the Glisson pedicle of left hepatic was (3.1±0.76)cm. The length of the the Glisson pedicle of middle hepatic was (2.61±0.72)cm. The length of the the Glisson pedicle of right hepatic was (1.5±0.50)cm. The shapes of the Glissonean pedicle stem of the left hepatic presenting arch, the number of radial level 3 branches were between 2-8. The shapes of the Glissonean pedicle stem of the middle hepatic continuing the main of Glissonean pedicle, the number of radial level 3 branches were between 2-6. The shapes of 30% of the Glissonean pedicle of the right hepatic presenting Y and V, 70% of the Glissonean pedicle of the right hepatic presenting C, the number of radial level 3 branches were between 3-8. Conclusion In the light of morphology,embryology and comparative anatomy, it is reasonable to divide the liver into left,middle,right lobe by Glissonean pedicle of radial level 2 branches and the liver is an axiality and symmetry organ. References | Related Articles | Metrics

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Anatomy and preliminary clinical applications of endoscopic endonasal approach to optic canal and orbit GE Jun-qi ZHANG Xiao-biao* HU Fan YU Yong GU Ye SUN Cong-jing 2014, 45 (5):  688-693.  doi: 10.3969/j.issn.0529-1356.2014.05.018 Abstract ( )   Objective To provide the endoscopic anatomic basis and anatomic parameters for endoscopic surgical therapy on orbital lesions, and to analyze the advantages and key points of this surgical approach. Methods Five fresh adult heads were used in this experiment. Endonasal thanslamina approach and endoscopic technique were applied to observe important anatomic marks while intraoperative medicalization of the medical rectus muscle was applied to observe the exposure and positioning of important structures and trends of the optic canal and intra orbit. Results Uncinate process was at the lower front corner of middle nasal concha; ethmoidalis bulla was behind the uncinate process, and ethmoidei sinus was reachable after an incision was made on ethmoidalis bulla; anterior ethmoidal artery and posterior ethmoidal artery were the important anatomic landmarks of the inner ethmoidei sinus; optic canal prominence, carotid artery prominence and OCR were the important anatomic landmarks of the inner sphenoid sinus; lamina papyracea was at the lateral wall of ethmoidei sinus, and orbital contents were approacchable after lamina papyracea was cut off; inside orbit, the optic nerve was approachable through the gap between the medial rectus muscle and inferior rectus muscle. The ophthalmic artery of 9 out of 10 sides of the specimens was originated from the supraclinoidal segment of the internal carotid artery while the remaining one was originated from the cavernous segment of internal carotid artery. There were 7 sides in which ophthalmic artery was located at the inferior lateral of the optic nerve; there were 2 sides in which ophthalmic artery was located at the inferior of the optic nerve; the remaining one was located at the inferior medial of the optic nerve. Conclusion The endoscopic endonasal thanslamina approach can sufficiently expose the optic nerve and the structures in the medical space of the orbit. Uncinate process, ethmoid bulla, anterior ethmoidal artery, posterior ethmoidal artery and posterior ethmoid sinus are the important landmarks of the endoscopic endonasal thanslamina approach. Optic canal prominence，internal carotid artery prominence and OCR are the important landmarks for optic canal decompression. Ophthalmic artery, orbital branches, anterior ethmoidal artery, posterior ethmoidal artery, internal carotid artery are the important vessels. Medialization of the medial rectus muscle is effective to approach the orbital anatomical structures. References | Related Articles | Metrics

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Measurement of the femoral neck torsion angle and anteversion angle by laser projection method ZHU Qiu-liang* XU Bing SHEN Liang-hua ZHAO Li-lai YAN Mao-hua WANG Yuan-long ZHANG Ying 2014, 45 (5):  694-697.  doi: 10.3969/j.issn.0529-1356.2014.05.019 Abstract ( )   Objective To investigate the feasibility of measuring femoral neck torsion angle and anteversion angle by laser projection method. Methods The femoral neck torsion angle and anteversion were observed and described. An angle measuring device was designed and produced. With the device, the femoral torsion angle and anteversion angle were measured by laser projection method two times. Statistical analysis was performed on the measured value, and sides difference. Results The differences between femoral neck torsion angle and anteversion angle were observed. There was no significant difference (P> 0.05, power=100%) between the two measurements by laser projection method. Measurements of the femoral anteversion were 13.58 °±6.55 ° on the left side, and 12.15 °±5.83 °on the right side. Measurements of the femoral neck torsion angle were 18.50 °±7.38 ° on the left and 19.08 °±8.59 ° on the right. There was no significant difference between left and right side (P > 0.05). Conclusion The laser projection method is the effective method in measuring femoral neck torsion angle and anteversion angle, and has excellent repeatability. References | Related Articles | Metrics

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Ventricularization of the proximal cardiac outflow tract contributes to trabeculated right ventricle in mouse embryo LI Hai-rong YANG Yan-ping WANG Jing-jing CAO Xi-mei LIU Hui-xia CUI Hui-lin ZHANG Tao JING Ya* 2014, 45 (5):  698-703.  doi: 10.3969/j.issn.0529-1356.2014.05.020 Abstract ( )   Objective To explore the mechanism underlying the rapid shortening of outflow tract and the formation of the right ventricle of the embryonic mouse heart. Methods Serial sections of embryonic mouse hearts from embryonic day 9 (E9) to E12（3 to 5 embryos for each stage）were stained with antibodies against α-sarcomeric actin (SCA), α-smooth muscle actin (SMA), GATA-4, myosin heavy chain (MHC), proliferating cell nuclear antigen (PCNA) or active caspase-3 (CAS-3). Results At E11, the aortic sac and the distal border of cardiac outflow tract had regressed towards the ventricle into the pericardial cavity, while GATA-4、SCA and SMA staining showed that precursors from the second heart field were differentiating into cardiomyocytes adding to the arterial pole of the heart to lengthen the outflow tract. The length of outflow tract rapidly shortened at E12. Before and during its shortening, no CAS-3 positive cell was detected in the entire outflow tract. During E10-12, the cardiomyocytes in the right ventricle and proximal outflow tract wall proliferated inward to form trabeculae, with some trabeculae extending into the ridges. Proximal extremities of the outflow tract ridges were gradually myocardialized remodeling into the trabeullar right ventricle wall. At E12, scattered SCA and SMA staining cells and SCA and SMA weak positive mesenchymal cell clusters, which were continuous with the outflow tract myocardium were detected in the mesenchymal proximal outflow tract ridges. These results suggested that the proximal outflow tract was remodeled into the right ventricle by trabecularization, during which mesenchymal ridges were trabecularlly myocardialized. Conclusion Ventricularization of the proximal outflow tract contributes to the trabecular right ventricle and resultes in the vapid shortening of outflow tract in the mouse embryonic heart. Cardiomyocyte appoptosis and transdifferentiation are found to play a more limited contribution during this process. References | Related Articles | Metrics

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Cadmium damages the blood-testis barrier in rats and the protective effect of Astragaloside IV NING Wei LIAO Xiao-gang* WANG Yi YAO Zhi-yong MAO Sheng-yan 2014, 45 (5):  704-709.  doi: 10.3969/j.issn.0529-1356.2014.05.021 Abstract ( )   Objective To observe the effect of astragaloside IV (A) and SB203580 antagonize cadmium(Cd) toxicity on expression of associated protein and blood-testis barrier(BTB) in rats and to study the protective mechanism of A on it. Methods Totally 21 SD male rats were randomly divided into 7 groups, 3 rats per group：Cd ［intraperitoneally injected with 0.1%CdCl2，1mg/(kg·d)］，Cd+A ［at the above dose of CdCl2，at the same time with A，10mg/(kg·d)］，and Cd + SB203580 ［at the above dose of CdCl2，at the same time with SB203580，100μg/(kg·d)］， each of the above groups was further divided into continuous five and ten days treatment groups. The control group was intraperitoneally injected with equal dosage of normal saline. The testes were studied by light, electron microscopy, immunohistochemistry and Western blotting. Results In the control group，irregular and lightly stained nuclei of Sertoli cell(Sc) in seminiferous tubules were observed by HE staining. A continuous electron density line of tight junction（TJ）and normal ultrastructure of BTB were observed. After Cd treatment，the vesicular formation in the Sc was observed. The ultrastructural damage of Sc and TJ was observed. Compared with the corresponding time point of Cd group，these were weakened in morphology of testis and ultrastructure of TJ after Cd + A or Cd + SB203580 treatment. The positive products of zonula occludens-1（ZO-1） and claudin-11 were localized mainly in the base of the seminiferous tubule. After Cd treatment，the average absorbance (AA) of ZO-1 and Claudin-11 was decreased significantly compared with that of the control group（P＜0.05）. After Cd + A or Cd + SB203580 treatment，AAof ZO-1 and Claudin-11 were increased significantly compared with that of the Cd group（P＜0.05），though lower than that of the control group. The result of Western blotting showed that phosphorylation-p38MAPK in Cd group was increased significantly compared with that of the control group（P＜0.05）.After Cd + A or Cd + SB203580 treatment, it was decreased significantly compared with that of the Cd group（P＜0.05）. Conclusion Cd decreases ZO-1 and Claudin-11 expression and damages ultrastructure of TJ in BTB, as Ⅳ has protective effect on it, and is related to inhibiting activation of p38 MAPK pathway. References | Related Articles | Metrics

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Association between body composition and blood lipids in pre-and post-menopausal women of Maonan ethnicity DENG Qiong-ying* JIANG Xian-yong YU Hong-rong ZHOU Li-ning GONG Ji-chun DENG Qiu-yun 2014, 45 (5):  710-714.  doi: 10.3969/j.issn.0529-1356.2014.05.022 Abstract ( )   Objective To study the differences in body composition and blood lipids between the pre-and post-menopausal women of Maonan ethnicity, and to explore the correlations between body fat content, fat distribution and blood lipids. Methods Totally 200 Maonan pre-and post-menopausal women were randomly selected from Huanjiang county in Guangxi. Body composition were measured by bioelectrical impedance analysis(BIA), and blood lipids were tested from blood samples. Results Compared with the pre-menopausal women, the visceral fat level (/area), waist-hip ratio(WHR), left (/right) lower limbs fat, total cholesterol (TC), triglyceride (TG) and low-density lipoprotein( LDL-C) in post-menopausal women were significantly higher (P<0.01), and the detection rate of hypercholesterolemia, mixed hyperlipidemia and dyslipidemia in postmenopausal group was also significantly higher(P<0.01). All the blood lipids were closely related to WHR and visceral fat content (P<0.05 or P<0.01). In addition, TG, high-density lipoprotein (HDL-C) and LDL-C except TC were significantly correlated to %BF, BMI and subcutaneous fat content (P<0.05 or P<0.01). Conclusion The accumulation of visceral and abdominal fat in Maonan postmenopausal women is significantly correlated to dyslipidemia. The results may provide references for making preventive program for the Maonan women. References | Related Articles | Metrics

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Somatotype characteristics of Bai ethnicity children and adolescents in Hu’nan HUANG Da-yuan ZHANG Hui-juan WU Guo-yun LIANG Cheng-qing ZHU Yao-feng 2014, 45 (5):  715-718.  doi: 10.3969/j.issn.0529-1356.2014.05.023 Abstract ( )   Objective To study the characteristics and regularities of somatotype of Bai ethnicitty children and adolescents in Hu’nan. Methods The somatotype growth of 1525 Bai children and adolescents (male: 748, female: 777) was evaluated by the Heath-Carter anthropometric method. Results The mesomorphy of male was bigger than that of female, and endomorphy of female was bigger than that of male. The primary somatotype in male was mesomorph and ectomorph, and it was central and ectomorph in female. The somatotypes developed from balanced mesomorph, ectomorphic mesomorph, mesomorph-ectomorph, mesomorphic ectomorph to mesomorph-ectomorph in male; however, in female from central, balanced ectomorph, central, endomorph-mesomorph to mesomorphic endomorph. Conclusion The somatotypes are very different between males and females of Bai ethnicity children and adolescents. The somatotype of males is slender with less fat and more muscular, however, they are plumper with more body fat and shorter stature in females. Bai ethnicity children and adolescents have less muscular, less fat and shorter stature than Mongolian and other populations. References | Related Articles | Metrics

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Physical traits of the body of Chishui Miao ethnicity in Guizhou YU Xiao-dan* TANG Li-jun LI Feng-hua JIANG Gui-chuan CHEN Kai-qin WU Shi-rong 2014, 45 (5):  719-723.  doi: 10.3969/j.issn.0529-1356.2014.05.024 Abstract ( )   Objective To accumulate data for the physical anthropology research of Miao ethnicity adults, and find out the kinship and difference between this group and the other 10 ethnicities. Methods Viviperception and measurement were used to study the caudomedial part traits in 299 Miao ethnicity adults (146 males and 153 females ) who lived in Chishui city in Guizhou, and statistical software SPSS18.0 was used to process data. Results Apart from length of middle finger, the height of medial malleolus subpoint, and the rest 19 indices between male and female had significant difference or great significant difference (0.01＜P＜0.05orP＜0.05). Great majority of males were trunk types (55.5%) and more females tended to be the long trunk type (39.9%). Most of males belonged to wide chirality (44.40%) while more females tended to be narrow chirality (39.9%). More males were submakroskelic type (33.6%) and majority of females tended to be mesatiskelic type (35.3%). Males and females tended to the hypermicrosoma (males 37.0%, females 54.9%). Conclusion The physical character of the Miao ethnicity males and females living in Chishui city is positively related to the Miao ethnicity in Xishui. There is significant difference between Miao and Mulao ethnicity. References | Related Articles | Metrics

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A new method to study nerve fiber projecting in the spinal cord during chicken embryo development HU A-zhen1 YANG Ci-qing FU SU-lei JIA Yang-yang LI Han GUO Zhi-kun LIN Jun-tang * 2014, 45 (5):  724-728.  doi: 10.3969/j.issn.0529-1356.2014.05.025 Abstract ( )   Objective To develop a method of studying fiber projecting in the spinal cord duiring chicken embryo development. Methods At embryonic incubation 3 day (E3), pCAGGS-green fluorescent protein (GFP) plasmid was injected into the spinal cord usingin vivo electroporation. Three days after transfection (E6), GFP-positive embryos were collected under a stereo fluorescence microscope. Subsequently, the spinal cord was separated from the embryos and cut from the roof plate as an open book. After fixed with 4% paraformaldehyde (PFA) for one hour, the opened spinal cords were used for immunohistochemistry with N-cadherin antibody and with DAPI for nuclei. Finally, the nerve fiber projecting was photographed and analyzed under a fluorescence microscope. Results Based on the opened spinal cord and immunostaining in the cryosection, we observed that the nerve fibers projected across the midline of the floor plate and reached to the sulcus terminalis along the white matter of the contra side. The immunoreaction against N-cadherin indicated that overexpression of GFP has no significant effect on chicken embryonic development. Conclusion A new method to study fiber projecting in the developing chicken spinal cord is established successfully in this study. References | Related Articles | Metrics