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2014, Volume 45 Issue 2
06 April 2014 Previous Issue    Next Issue

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Molecular mechanism of the chronic alcohol exposure induced injury of the liver-brain in SMS2 knockout mice ZHAO Jing-ya CUI Zhan-jun HE Wei-ya SUN Yu-hua WANG Si-qian DENG Jin-bo* LU Guang-xiu* 2014, (2):  145-154.  doi: 10.3969/j.issn.0529-1356.2014.02.001 Abstract ( )   Objective To investigate the relationships among the chronic alcohol exposure, stress injury of liver-brain and ceramide’s effect. Methods The chronic alcohol exposure models in both wide type (WT) and sphingomyelin synthetase 2 knockout (SMS2-/-) mice were established. HE staining, Masson staining, c-Fos and NF-κB immunolabeling, Western blotting analysis and trranssmission electron microscopy were carried out. Results Alcohol exposure led to the wild-type and SMS2-/- mice hepatocytic steatosis and hepatic fibrosis. Mallory bodies and inflammatory cell infiltration were found in liver. After alcohol exposure, c-Fos and NF-κB positive cells increased in hippocampal CA1 area with dose dependency (P<0.01). However, comparing with the WT pups, the number of positive c-Fos and NF-κB cells in SMS2-/- mice increased much more than WT mice (P<0.05). Immunoblotting evidence of c-Fos and NF-κB protein supported the data of immunohistochemistry in both in WT and SMS2-/-mice (P<0.01 )Conclusion Oxidative stress and inflammatory responses in liver and CNS are the main pathological alterations after chronic alcohol exposure. The alcoholinducing oxidative stress probably is the cause of insulin resistance in organism. Ceramide is involved in the processes above through c-Fos and NF-κB pathway. Alcohol exposure induces the oxidative stress injury in liverbrain axis, such as inflammatory responses and the expression of oxidative stress cues, c-Fos and NF-κB. Ceramide increases the alterations above. References | Related Articles | Metrics

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Vascular and stem-cellular niche and the neural proliferation and differentiation in spinal cord of the mouse XI Yan XUE Shuai LI Li-li NIE Meng-yue DENG Jin-bo* 2014, (2):  155-160.  doi: 10.3969/j.issn.0529-1356.2014.02.002 Abstract ( )   Objective To study how the vascular and stem-cellular niche affect the neural proliferation, differentiation and migration in the mouse spinal cord and to investigate the interaction among neurons, neuroglia and vasculature in the developing spinal cord. Methods A total of 150 mice from E17 to P180 were used in this project. BrdU assay, Nissl’s staining and immunofluorescent labeling (NeuN and GFAP) methods were applied. Blood vessels and neural stem cells were measured with stereological methods. Results BrdU positive neural stem cells appeared evenly in the embryonic spinal cord at as early as E14, and the newborn neurons migrated from the subventicular zone of the neural tube to the intermediate layer around it. With age increasing, the neural stem cells differentiated into neurons gradually, and the neurons mainly concentrated in the putative gray matter as a “H” shape. According to the statistical analysis, the BrdU positive stem cells developed as parabolic curve with the peak at P3. The neuroglial cells migrated from the “periphery” area of spinal cord (marginal layer, the putative white matter) into the “center” (gray matter). Blood vessels were uniform distribution at the early age, and later concentrated in the gray matter gradually. The vasculature, stem cells, neurons and neuroglia formed a so called vascular and stem-cellular niche. The vascular stem-cellular niches were mainly located in the subventricular zone and gray matter. Vasculature, stem cells, neuron and neuroglia were woven together tightly. Conclusion The vascular and stem-cellular niche formed closely with the development of the spinal cord, which provides a suitable microenvironment for the development of the spinal cord and the differentiation of neurons and neuroglia. References | Related Articles | Metrics

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Silencing AQP4 gene in the astrocytes through siRNA transfection by lipofectamine RNAiMAX ZHANG Wei XU Li-xin YANG Shao-hua SHI Zhong-fang DONG Li-ping YUAN Fang* 2014, (2):  161-165.  doi: 10.3969/j.issn.0529-1356.2014.02.003 Abstract ( )   Objective To determine whether siRNA transfection with the help of Lipofectamine RNAiMAX can achieve AQP4 gene silencing in primary cultured astrocytes. Methods Primary cultured astrocytes from Wistar rat’s cerebral cortex were used in this study. Fluorescence microscope and Tecan microplate reader were used to examine whether Cy3-labeled siRNA can be transfected into astrocytes by lipofectamine RNAiMAX and the effects of different RNAiMAX concentration and siRNA concentration on the transfection efficiency. The silencing effect of AQP4 gene was detected by Real-time PCR. Results There were a lot of transfected cells under different transfection conditions as observed by fluoscence microscope. Tecan micropalte reader detection showed that the fluorescence intensity enhanced when siRNA and RNAiMAX concentrations increased. The level of AQP4 mRNA decreased more than 80 percent at 24 hours, 48 hours and 72 hours after 3ml/L RNAiMAX and 30nmol/L AQP4 siRNA or 6ml/L RNAiMAX and 60nmol/L AQP4 siRNA were used to transfection observed by Real-time PCR(n=6). Conclusion The fact that AQP4 mRNA significantly decreased after transfection indicates that siRNA can be successfully transfected into primary cultured astrocytes and silenced AQP4 gene by using Lipofectamine RNAiMAX. References | Related Articles | Metrics

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Distribution of gonadotropin releasing hormone receptor in cervicothoracic ganglion of female goats DONG Wei LI Qiang XU Yong-ping* JIN Xiu-fang CHEN Wen-dong GUO Xiao 2014, (2):  166-170.  doi: 10.3969/j.issn.0529-1356.2014.02.004 Abstract ( )   Objective To explore whether cervicothoracic ganglion has the condition by which gonadotropin releasing hormon(GnRH) can exert its function. Methods Immunohistochemical SP method was adopted to observe the distribution of GnRH receptor (GnRHR) in cervicothoracic ganglion of female goats. Results The results showed that, all the neurons in cervicothoracic ganglion responded positively to GnRHR. The membrane of neurons and the granular material in cytoplasm were strong positive. The neuronal nucleus was negative to GnRHR while neurites and satellite cells showed medium positive. Image analysis showed a significant difference of GnRHR expression between neurons and non-neurons (P＜0.01).Conclusion The results indicate that neurons in cervicothoracic ganglion are the main target cells of GnRH, and cervicothoracic ganglion has the condition by which GnRH can exert its function, which implies GnRH may regulate the function of heart and lung through the GnRHR distributed in cervicothoracic ganglion. Thus，the GnRHR existing in these neurons may be a “transfer station” for GnRH to coordinate heart and lung activity through endocrine and autonomic nerve pattern. References | Related Articles | Metrics

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Distribution characteristics of follicle-stimulating hormone receptor in the cervicothoracic ganglion of goat JIN Xiu-fang WANG Zhi-hao FAN Jie XU Yong-ping* CHEN Wen-dong GUO Xiao 2014, (2):  171-175.  doi: 10.3969/j.issn.0529-1356.2014.02.005 Abstract ( )   Objective To investigate whether follicle stimulating hormone (FSH) affects autonomic regulation of cardiopulmonary function. Methods Five pairs of cervicothoracic ganglion were taken from mature female and male gosts. The distribution of follicle stimulating hormone receptor (FSHR) in the cervicothoracic ganglion of the goat was observed after immunohisto-chemical SP staining. Results FSHR immunoreactive substances mainly distributed in neurons’ cytoplasm and membrane, but not in the nucleus. There was some expression of FSHR in surrounding supporting cells, passing fibers,Schwan cells and vascular endothelial cells. Image analysis showed that the relative expression of immunoreactive product FSHR was significantly higher than that of other nonneural structures(P＜0.01). Conclusion These results suggest that the neurons in cerricothoracic ganglion are the main target cells of FSH in goats and FSH is likely to affect neurons of the cervicothoracic ganglion and then further to regulate cardiopulmonary function activities via the postganglionic sympathetic nerve. References | Related Articles | Metrics

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Expression and significance of Nestin and SOX2 in the SAMP8 mice subfornical organ YE Jian-ya LI Cong-hui TANG Wei-dong MA Chang-sheng HAO Qing-mao QIN Yong* 2014, (2):  176-180.  doi: 10.3969/j.issn.0529-1356.2014.02.006 Abstract ( )   Objective To observe the expression of nestin and sex-determining genes in high-mobility group proteins in the senescence accelerated mouse prone 8 subfornical organ. Methods Adult SAMP8 mice (8 months old) were used. The expressions of Nestin, SOX2 in subfornical organ were observed by using immunohistochemical staining and immunofluorescence staining methods. Normal senescence mouse resistance 1 was displayed as control. Results Nestin immunohistochemical staining of the control group: positive cells were protuberantly distributed and expressed filiform texture with radiate. Nestin immunohistochemical staining of the experimental group: positive cells body stained stronger, filiform texture was thick, densely distributed in surrounded by vessels. The percentage of positive cells（49.17±7.60）% increased significantly than control group（16.33±4.41）% and had statistical significance (P<0.01). SOX2 immunohistochemical staining of control group: positive cells stained irregularity and dispersedly distributed. SOX2 immunohistochemical staining of the experimental group: most of positive cells stained deeper, densely distributed in surrounded by vessels. The percentage of positive cells（62.17±20.27）% increased significantly than control group（36.00±16.20）% and had statistical significance (P<0.05). Fluorescence double staining in the control group: SOX2 stained irregularity and Nestin expressed during it. A small amount of SOX2 and Nestin double-positive cells were in the subependymal region. In the experimental group: positive cells expressed intensity and showed more double-positive cells. Conclusion Expression of nestin and SOX2 enhances significantly in the senescence accelerated mouse prone 8 subfornical organ, which indicates AD may result in increased number of neural stem/progenitor cell proliferation or differentiation in subfornical organ at same stage，and then the neurogenesis of subfornical organ may be affected in Alzheimer disease. References | Related Articles | Metrics

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Morphological changes and NOS expression of neurons in CA3 hippocampus of rats LIU Ji*WANG Xiao-yu SUN Yang GUO Xiu-ying PANG Yin 2014, (2):  181-184.  doi: 10.3969/j.issn.0529-1356.2014.02.007 Abstract ( )   Objective To observe morphological changes and the nitric oxide synthase (NOS) expression of neurons in CA3 hippocampus of rats and investigate morphological changes of hippocampal neurons and the role of nitric oxide (NO) in the pathogenesis of hepatic encephalopathy. Methods Fifty male Wistar rats were divided into the control group and experimental group. Before the experiment all the rats were tested by Morris water maze test. After 9 weeks, a CCL4 model of hepatic encephalopathy was established. The hippocampus of each group was taken out for Nissl staining and dyeing of NADPH-d. Results 1.Nissl’s staining: In the experimental group, the number of neurons in hippocampus were less than the control group and dyed lighter, Nissl bodies were also reduced. 2. NADPH-d staining results: In the experimental group, the axon was dyed more deep，widespread dendrites connections were observed. In the control group there were a few positive axons and less connections between dendrites. The color in the experimental group was blue or deep blue and showed positive or strong positive with NADPH-d staining, while in the control group showed weak positive. Conclusion Hepatic encephalopathy can lead to hippocampus injury. NO may mediate the process and the blood ammonia level is one of the pathogenesis of HE. References | Related Articles | Metrics

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Stereoscopic study on the expression of Bcl-2 and Bax in development of mice hippocampus ZHANG Li ZHANG Jing-kun GUO Min* 2014, (2):  185-189.  doi: 10.3969/j.issn.0529-1356.2014.02.008 Abstract ( )   Objective To observe the expression of Bcl-2 and Bax in the development of C57/BL6 mice hippocampus. Methods Mice hippocampus of Embryos (E) 18 day (d), E 20 d, postnatal (P) 1 d, P 3 d, P 7 d, P 14 d, P 28 d, P 2 month (M), P 3 M, P 6 M, P 15 M, P 18 M were used in this study. Immunohistochemistry and stereoscopy were used to detect the expression of Bcl-2 and Bax protein in the mice hippocampus at different ages. Results From E18 d to P21 d, Bcl-2 and Bax immunoreactive cells in both Ammon’cornu (CA) and dentate gyrus (DG) firstly and gradually increased and then decreased（P<0.01）. The volume density of both Bcl-2 and Bax immunoreactive cells came to the top on P7 d except the Bax expression in DG which was the highest on P14 d（P<0.01）. Conclusion The expression of Bcl-2 and Bax is high at the early stage of hippocampus development and then low at its adult stage and senectitude, which may be involved with the forming of hippocampus. References | Related Articles | Metrics

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Effect of XAV939 on proliferation inhibition of human neuroblastoma cell TIAN Xiao-hong HOU Wei-jian BAI Shu-ling* FAN Jun TONG Hao XU He 2014, (2):  190-195.  doi: 10.3969/j.issn.0529-1356.2014.02.009 Abstract ( )   Objective To study the effect of antiproliferative and apoptosis induction of small molecule inhibitor XAV939 on neuroblastoma (NB) cell line SH-SY5Y cells, and its possible mechanism. Methods The MTT assay was used to detect the inhibition effect of XAV939 on the viability of SH-SY5Y cells, and to determine the optimal concentration. Annexin V-FITC assay was used to detect the percentage of early apoptotic cells after XAV939 treatment. Hoechst33342 staining was used for observing the apoptotic nuclear morphology. Colony forming assay was applied for detecting the proliferation effect of XAV939 on SH-SY5Y cells. The expression of key proteins of Wnt/β-catenin pathway and anti-apoptotic protein Bcl-2 were detected by Western blotting. Results The MTT assay showed that the proliferation inhibition rate of SH-SY5Y cells increased significantly by 1μmol/L XAV939 for 24hours. The percentages of apoptotic cells were significantly higher than those in control groups after treated with XAV939 for 48hours and 72hours, and the cells showed varying degrees of apoptotic morphology. In addition, XAV939 significantly reduced the colony formation rate of SH-SY5Y cells in vitro. The results of Western blotting showed that XAV939 reduced the expression of key proteins of Wnt/β-catenin signaling pathway as well as anti-apoptotic protein Bcl-2. Conclusion XAV939 may inhibit the proliferation of SH-SY5Y cells in part by inhibiting the Wnt/β-catenin signaling pathway. References | Related Articles | Metrics

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Down-regulation of tankyrase 1 mediated by RNAi leads to the apoptosis of SH-SY5Y cells TIAN Xiao-hong BAI Shu-ling* FAN Jun HOU Wei-jian TONG Hao XU He 2014, (2):  196-203.  doi: 10.3969/j.issn.0529-1356.2014.02.010 Abstract ( )   Objective To investigate the effect and mechanism of tankyrase 1（TNKS1）down-regulation mediated by RNA interference（RNAi）on the proliferation and apoptosis of human neuroblastoma (NB) SH-SY5Y cells, and to provide experimental basis for the development of a new therapeutical target. Methods According to TNKS1 gene sequences, three short hairpin (shRNA) interference segments with gene-specific were designed and synthesized, and lentiviral vectors were used for constructing RNAi sequence. After SH-SY5Y cells were transfected with lentiviral vector, Real-time PCR assay was used for detecting the expression of TNKS1 gene, and the best interference sequence was screened out for subsequent experiments. The colony forming assay was used to detect the change of cell proliferation after RNAi-TNKS1. The expression of anti-apoptotic protein Bcl-2, pro-apoptotic protein Caspase-3, the key protein β-catenin of Wnt/β-catenin pathway and its downstream target proteins Cyclin D1 and c-Myc were detected by the Western blotting method for exploring the mechanism. Apoptotic morphology was observed by transmission electron microscopy. Results The lentiviral vector was constructed successfully, and the virus concentration was 5×10 11TU/L. The results of Real-time PCR test showed that #3 shRNA was the most effective target sequence. The colony forming assay results showed that colony forming rate decreased significantly after RNAi-TNKS1 compared with that of control group. The results of Western blotting showed that the protein expression of Bcl-2 reduced significantly as well as β-catenin, Cyclin D1 and c-Myc after RNAi-TNKS1, while the protein expression of caspase-3 increased. The transmission electron microscopy results showed significant apoptotic structure and morphology after RNAi-TNKS1. Conclusion The down-regulation of TNKS1 mediated by RNAi leads to the proliferation decrease and the apoptosis of SH-SY5Y cells, and the inhibition of Wnt/β-catenin pathway may play a role in it. References | Related Articles | Metrics

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Identification of miRNAs regulating pancreatic duodenal homeobox gene 1 expression during the induction of bone marrow mesenchymal stem cells differentiation into insulin-producing cells WANG Tao MA Yun-sheng MU Chang-zheng* 2014, (2):  204-210.  doi: 10.3969/j.issn.0529-1356.2014.02.011 Abstract ( )   Objective To identify the miRNAs which may regulate the expression of pancreatic duodenal homeobox gene 1(PDX-1) during the induction of bone marrow mesenchymal stem cells(BMSCs) differentiation into insulinproducing cells. Methods BMSCs were isolated from bone marrow and induced to differentiate into insulin-producing cells using conophylline and nicotinamide. The miRNAs which may regulate the expression of PDX-1 were predicted using miRanda and TargetScan and identified by dual luciferase report system. The expressions of miRNAs and PDX-1 were determined using Real-time PCR during the induction of BMSCs differentiation into insulin-producing cells. Results The induced cells were stained scarlet by DTZ and the expression of insulin was positive by immunofluorescence cytochemistry. Four miRNAs which may regulate the expression of PDX-1 were firstly obtained by bioinformatics.Using dual luciferase reporter system in vitro, two of the four miRNAs, miR-149and miR-346, were proved to effectively inhibit PDX-1 expression, respectively, by binding to the 3’UTR of PDX-1 mRNAs. The Real-time PCR results showed that the expression of miR-149 or miR-346 was negatively correlated with that of PDX-1 mRNA. Conclusion The results indicated that miR-149 and mi-346 may negatively regulate the expression of PDX-1 during the induction of insulin-producing cells. References | Related Articles | Metrics

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Temporal expressions of fibroblast growth factor receptor genes during Zebrafish embryonic development ZHANG Jia-jia LIN Lin1 LIU Ni-ya WU Hong-mei HE Jin-cai* HUANG Chen-ping* 2014, (2):  211-215.  doi: 10.3969/j.issn.0529-1356.2014.02.012 Abstract ( )   Objective To study the temporal expressions of fibroblast growth factor (FGF) receptor genes (fgfrs) during zebrafish embryonic development. Methods Total RNA was extracted from zebrafish embryos at 4, 6, 10, 12, 18, 24, 48 and 72 hours post fertilization (hpf), respectively, and was reversely transcripted to cDNA. The fgfrs’ mRNA levels were then determined by real time quantitative polymerase chain reaction (Real-time PCR). Results The temporal mRNA expression of fgfrs varied significantly during embryonic development with a pattern of peak curve (P<0.01). During zebrafish embryogenesis the mRNA expressions of fgfr1a-b, fgfr4, fgfr2 and fgfr3 began at blastula, early gastrula, gastrula and late gastrula, and peaked at gastrula, late gastrula, segmentation and mid-late period, respectively. There was significant correlation between the expression levels of the paralogs fgfr1a and fgfr1b (r=0.830,P<0.05). And there were also significant correlations in fgfr1a-fgfr3, fgfr1b-fgfr4 and fgfr2-fgfr3 (P<0.05) with r values of -0.726, 0.821 and 0.772, respectively. Conclusion Each of fgfrs has its own expression pattern during zebrafish embryonic development. The fgfr1 and fgfr4 mainly play their roles at the early period of embryogenesis, while the fgfr2 and fgfr3 at the mid-late period. The co-expression of fgfr1a and fgfr1b exhibits the redundant function of the paralogs during embryonic development. There probably is a reciprocal relationship in gene expression between fgfr1 and fgfr3, But the intrinsic correlations of fgfr1-fgfr4 and fgfr2-fgfr3 still need to be verified. References | Related Articles | Metrics

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Effects of the uremic toxin p-cresylsulfate on human umbilical vein endotheilial cell function in vitro LIN Wei* XIA Jian-lan WANG Yi 2014, (2):  216-220.  doi: 10.3969/j.issn.0529-1356.2014.02.013 Abstract ( )   Objective To investigate the effect of a uremic toxin, p-cresylsulfate, on human umbilical vein endothelial cells (HUVECs) function. Methods HUVECs were incubated with various concentrations of p-cresylsulfate (10, 40, 80, and 160 mg/L) for up to 72 hours. Cell proliferation was determined using WST-1 assay. Tube formation assay was used to evaluate HUVECs function in vitro. Reactive oxygen species (ROS) production in HUVECs was measured by fluorescence microscopy. Inflammatory factor and adhesion molecule expressions were assessed by real time quantitative PCR(Real-time PCR) and Western blotting analysis. Results P-cresylsulfate had no anti-proliferative effects on HUVECs. Cells treated with concentrations of p-cresylsulfate in the range found in CKD patients had no observable impairment on tube formation. In contrast, p-cresylsulfate partially increased ROS production in HUVECs, while quantitative Real-time PCR and Western blotting revealed p-cresylsulfate treatment increase interleukin(IL)-1β, tumor necrosis factor(TNF-a), and intercelluar adhesion molecule\|1(ICAM-1) expressions in HUVECs. Conclusion Our study revealed that a uremic toxin, p-cresylsulfate, have pro-oxidant and pro-inflammatory effects on HUVECs, which indicates that p-cresylsulfate may be among the factors involved in the high incidence of atherosclerosis in patients with chronic kidney disease. References | Related Articles | Metrics

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Expression and localization changes of nucleophosmin during apoptosis of human esophageal cancer EC9706 cells induced by curcumin SHI Song-lin CHEN Lan-ying LIU Yong-jin YANG Hai-bo LU Kun YANG Ling LI Qi-fu* 2014, (2):  221-227.  doi: 10.3969/j.issn.0529-1356.2014.02.014 Abstract ( )   bjective To explore the expression and localization change of nucleophosmin(NPM) in the nuclear matrix of cancer cells and its relationships with apoptosis regulating proteins during induced apoptosis, and to discuss its regulating functions in apoptosis. Methods Curcumin was used to induce apoptosis of human esophageal cancer EC9706 cells. By using subcellular proteomics technology we analyzed the presence and expression change of NPM in the nuclear matrix. We performed a Western blotting analysis to confirm the change. Subsequently, we examined the localization change of NPM and its co-localization with apoptotic gene products such as Bax and Bcl-2 through laser scanning confocal microscopy. Results The results of this study showed that NPM existed in the nuclear matrix of EC9706 cells with a remarkable down-regulation after curcumin treatment. We demonstrated that NPM exhibited a distinct nucleocytoplasmic shuttling during induced apoptosis. NPM was co-localized with Bax, Bcl-2 and their colocalization regions changed during the process of apoptosis. Conclusion NPM is a nuclear matrix associated protein. Its altered expression and localization, as well as its co-localization with apoptosis regulating proteins suggest the important regulative functions in apoptosis of EC9706 cells. References | Related Articles | Metrics

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Expressions and implications of end-binding protein 1 in autophagic Siha cells LIU Yi-qun LUO Xiong-yan JIANG Hong XIA Yan-hui LIU Qing-song* 2014, (2):  228-233.  doi: 10.3969/j.issn.0529-1356.2014.02.015 Abstract ( )   Objective To explore the expressions and implications of end-binding protein 1(EB1) in autophagic cervical cancer Siha cells. Method Real-time quantitative PCR and Western blotting were introduced to detect the mRNA and protein expressions of EB1, LC3 and p62 in Siha cells treated with Hank’s balanced salt solution (HBSS) or colchicinein vitro. Specific green fluoresent protein(GFP)-LC3 was then be detected to confirm the autophagosome formation by fluorescence microscopy withFlowCellectTMGFP-LC3 reporter autophagy assay kit. Result The mRNA and protein levels of LC3 and EB1 in cervical cancer Siha cells were increased significantly when prolong the duration of HBSS treatment (P<0.05). EB1 mRNA was positively correlated with LC3 mRNA expression (P<0.05). The expressions of p62 mRNA and protein reduced dramatically after HBSS treated (P<0.05), and was negatively related with the EB1 mRNA (P<0.05). The number of cells with GFP-LC3 and intensity of GFP-LC3 per cells were increased when starvation for 12 hours. However, the mRNA and protein levels of LC3, p62 and EB1 were not significantly changed when the cells treated with colchicine (P>0.05), and we cannot detected the fluoresce signaling of GFP-LC3 in cells at this moment. Conclusion The expression of EB1 in starved cells is dramatically increased, and accompanied with the increased expression of LC3 and autophagosome, decreased expression of p62. However, when microtubules with which EB1 interaction were destructed by colchicine, the above phenomenon disappears. All these results fully indicate the probably roles of EB1 in autophagy. References | Related Articles | Metrics

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Expression of galectin-3 in annexin A7 knockdown HeLa cells LI Xin* WANG Xiao-jie XU Qian CHEN Long LIANG Xiu-jun 2014, (2):  234-238.  doi: 10.3969/j.issn.0529-1356.2014.02.016 Abstract ( )   Objective To detect the expression of galectin-3（gal-3） in annexin A7 knockdown HeLa cells. Methods The HeLa cells were grouped into an experiment group, a negative group and a blank group. RNA interference technology was used to transfect the siRNA targeted to ANXA7, scrambled siRNA into HeLa cells with lipofectine 2000 without any treatment to the blank group. After transfection for about 48 hours, Western blotting method and PCR method were used to assure the suppression of ANXA7 on both protein level and mRNA level. The expression of galectin-3 was detected with Western blotting and real time qualitative PCR. Results The expression of ANXA7 was significantly suppressed in the cells transfected with ANXA7 siRNA. When ANXA7 expression was suppressed, the expression of galectin-3 decreased significantly compared with the negtive control group and blank control group. The results have significant meaning（P<0.05）. Conclusion The expression of galectin-3 is down-regulated in the ANXA7 suppressed HeLa cells, which may be one of the mechanisms for the abnormal behaviour of the ANXA7 knockdown carcinoma cells. References | Related Articles | Metrics

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Role of Notch signaling in platelet derived growth factor-BB induced proliferation of human pancreatic adenocarcinoma MA Yong-chao FAN Wen-juan WU Hua WANG Fu-qing* 2014, (2):  239-241.  doi: 10.3969/j.issn.0529-1356.2014.02.017 Abstract ( )   Objective To clarify Notch signaling in platelet derived growth factor-BB(PDGF-BB) induced proliferation of human pancreatic adenocarcinoma (HPAC). Methods After being treated by PDGF-BB and (or) γ-secretase inhibitor DAPT, changes of HPAC cell proliferation were detected by MTT assay; Alterations of Notch level in HPAC cells were measured by Western blotting assay; Variations of capability in cardiolipin synthetic lecithin(CSL) binding to the promoter of target gene hes-1 were identified by chromatin immunoprecipitation (CHIP) assay. Results PDGF-BB facilitates the proliferation of HPAC cells, upregulates Notch level and enhances the capability in CSL binding to the promoter of hes-1. DAPT and PDGFR inhibitor treatment suppresses and invalidates PDGF-BB effection. Conclusion Notch-1 signaling plays an essential role in the modulation of PDGF-BB-induced proliferation of HPAC cells. References | Related Articles | Metrics

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Distribution of arteries and nerves in human esophagus wall HUANG Xiao-yong LI Yan DENG Yi-bo HUANG Jian ZHU Lei, CHEN Dao-bang* 2014, (2):  242-247.  doi: 10.3969/j.issn.0529-1356.2014.02.018 Abstract ( )   Objective To explore the distribution characteristics and relationship between the arteries and nerves in human esophageal wall.Methods Eighteen cadavers (14 Childhood cadavers and adult 4 cadavers) were used in this study. The silicone vascular perfusion and remodified Sihler’s staining technique were combined to show the intravascular and intramural nerves of the human esophageal wall. Results The vascular disbribution of the cervical and upper thoracic esophagus was less than the lower thoracic and abdominal esophagus. The vascular distribution of the anterior wall was less than the posterior wall. The branches of the arteries formed anastomosis with each other. The nerve distribution of the human esophageal wall was more in thoracic esophagus, and less in abdominal esophagus. The nerve distributions were similar in thoracic and cervical esophagus. Some nerves in upper part of the cervical esophagus connected the with pharyngeal plexus nerve branches. The intramural nerves branches connected with each other, forming an anastomosis network. The distribution of the arteries in the human esophageal wall did not have clear accompany relationship with the nerves in the wall. Conclusion Both of arteries and nerves in human esophagus wall form anastomosis networks, but without clear accompany relationship. References | Related Articles | Metrics

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Measurement of the diameter of segmental and sub-segmental pulmonary arteries of the right inferior lobe by multislice spiral CT angiography CHEN Guang-ping* DONG Hai-na WANG Xiao-qing ZHENG Yu-feng XU Min ZHAO Zhong-wei 2014, (2):  248-252.  doi: 10.3969/j.issn.0529-1356.2014.02.019 Abstract ( )   Objective To measure the numerical value of segmental and sub-segmental pulmonary arteries of the right inferior lobe by multislice spiral CT angiography,to provide reference data for the imageology diagnosis of lung arteriopathy. Methods Totally 274 patients with no cardiac pulmonary diseases were performed with thoracic enhanced CT scans.The patients were divided into gender groups and three age groups of 20-40 year,41-60 year,and above 60 year.The diameter of segmental and sub-segmental pulmonary arteries of the right inferior lobe on the MIP images were observed, measured,compared and analyzed between the gender groups and among age groups. Results The numerical value of the diameter of segmental and sub-segmental pulmonary arteries of the right lung was accurately measured on the MIP images. There were significant differences of the diameter of segmental and sub-segmental pulmonary arteries of the right inferior lobe between the male and the female groups（P<0.05）.There were significant differences of the diameter of segmental and sub-segmental pulmonary arteries between the gender groups and among age groups（P<0.05）,especially between the age groups of 20-40 year and 61-85 year. Conclusion The imaging of multislice spiral CT can clearly display segmental and sub-segmental pulmonary artery of the right inferior lobe.It is reliable to measure the segmental and sub-segmental pulmonary arteries. References | Related Articles | Metrics

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Anatomical comparison of auditory organs between two species of bats in Yongzhou of Hu’nan province LIAO Yang* YAN Rong-ling LUO Ying LUO Jin-qiang CHEN Jie 2014, (2):  253-256.  doi: 10.3969/j.issn.0529-1356.2014.02.020 Abstract ( )   Objective To understand and compare the auditory organs ofScotomanes ornatus(SO) andPipistrellus abramus(PA) in Yongzhou of Hu’nan province.Methods The auditory organ components of 5 SO and 12 PA obtained through micro-dissection manipulation were measured and analyzed statistically. Results Complete tympanic membrane(SO: 9; PA: 19), malleus(SO: 10; PA: 18), incus(SO: 10; PA: 18), stapes(SO: 6; PA: 14) and cochlea(SO: 10; PA: 24) were isolated successfully. Characteristic parameters such as length of malleus’ helve, length of incus’ crus longum, height and bottom width of cochlea, area of tympanic membrane and stapes’ foot plate were measured, and significant difference existed in these characteristic parameters between two species. Amplification times of sound pressure in middle ear of the two species were 27.37 (SO) and 40.99 (PA), respectively, also showed significant difference. Conclusion Two species of bats shaped obviously different but matching their own living environment and acoustic behaviors auditory organs structure, although they are both belong to the same branch of vespertilionidae. References | Related Articles | Metrics

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Anatomy of the cerebral arterial circle and its appearance on magnetic resonance angiography LIU Jian ZHOU Bo-jiang* 2014, (2):  257-262.  doi: 10.3969/j.issn.0529-1356.2014.02.021 Abstract ( )   Objective To explore the use of three-dimensional time-of-flight (3D-TOF) magnetic resonance angiography of the cerebral arterial circle of the display case, and to provide morphological data for 3D visualization of arterial maps of the constructed basilar arterial circle. Methods Cerebral arterial circle was measured between artery angle and distance in 10 cadavers. The images of magnetic resonance angiography were got by 3D time-of-flight (3D-TOF) from 30 cases of normal human brain and were processed by Siemens Syngo B17 workstation, from medical imaging to analyze the cerebral arterial circle. Results Display rate of the anterior cerebral artery was 100%, bilateral arrangement. The mean distance of the artery circle was (1.06±0.23) cm, the mean angle with internal carotid artery was (63.13±9.72)°, and the mean angle of the anterior communicating artery was (107.95±57.70)°. Display rate of the internal carotid arterial was 100%, and the mean angle of the anterior cerebral artery was (63.13±9.72)°, with the mean angle of the posterior communicating artery was (123.20±10.37)°. Display rate of the posterior cerebral artery was 100%, the mean distance of the artery circle was (0.56±0.12) cm, the mean angle of the posterior communicating artery was (70.00±8.87)°, and the mean angle of the basilar artery was (123.95±11.43)°. Display rate of the posterior communicating artery was 100%, the mean distance of the artery circle was (1.06±0.17) cm, and the mean angle of internal carotid artery was (123.20±10.37)°, and the mean angle of the posterior cerebral artery was (70.00±8.87 )°, and the mean angle of the middle cerebral artery was (131.15±15.21)°. Display rate of the anterior communicating artery was 80%, the mean distance of the artery circle was (0.23±0.16) cm, and with the mean of the anterior cerebral artery angle was (107.95±57.70)°. Conclusion This study confirms the anatomical composition of the cerebral arterial circle, measured the distance between the cerebral arteries, and measured the angle between the artery circle within the artery. The results show that 3D-TOF method magnetic resonance angiography can clearly display the composition of the cerebral artery circle, and three-dimensional reconstruction techniques from different angles display the cerebral artery circle. References | Related Articles | Metrics

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Anatomy of the extended endoscopic endonasal transsphenoidal approach to the cavernous sinus LIANG Qiang LI Qiang DING Yong-zhong* XIE Min ZHOU Wang-ning DUAN Lei 2014, (2):  263-266.  doi: 10.3969/j.issn.0529-1356.2014.02.022 Abstract ( )   Objective To explore the endoscopic endonasal approach to cavernous sinus via extended transsphenoidal approach and establish the anatomic basis for endoscopic endonasal treatment. Methods Eight fresh adult cadavers were studied bilaterally (n =16). Zero and 30 degree rod lens rigid endoscope was used during dissection. After cadaver preparation, extended endoscopic endonasal approaches were performed to access the cavernous sinus. Results Endoscopic expanded nasal approach revealed the cavernous sinus structure and relevant anatomic landmarks were good for the anatomical localization. Conclusion The extended endoscopic endonasal transsphenoidal approach is the best surgical approach to the cavernous sinus. References | Related Articles | Metrics

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Programmed analysis of the method for the decellularization of intact rat kidney in vivo ZHAO Li-na YU Ya-ling LIN Ke-zhi SHAO Ying-kuan WANG Zhi-bin CHEN Jia-jun MEI Jin* TANG Mao-lin* 2014, (2):  267-272.  doi: 10.3969/j.issn.0529-1356.2014.02.023 Abstract ( )   Objective To provide the underlying data for a new ideal method that can keep the intact rat kidney bioscaffold. Methods Decellularized rat kidney scaffolds was prepared by treating with Triton X-100, SDS sequentially in vivo and each steps of the method was observed separately.Forty SD rats were assigned to four groups randomly with 10 rats group. Normal rat kidneys were treated as control group after heparinization (CG), and then kidneys were perfused with 500ml heparinized PBS as H group, heparinized PBS and 1% Triton X-100 sequentially as HT group,heparinized PBS and 1% Triton X-100 and 1% SDS sequentially as HTS group. After each step of decellularization, the genomic DNA contect analysis, transmission electron microscopy, HE, Masson’s, PAS staining and immunohistochemistry techniques were used in order to observe changes in extracellular matrix constitution. Results Rat kidneys were decellularization in 6 hours. Its’cells and debris were cleaned gradually in the perfusion process and become translucent eventually. HE, Masson’s, PAS staining and transmission electron microscopy showed that the normal rat kidney was changed into reticular structure, it appeared to be continuous distribution and morphological arrangement was similar to the glomerular and tubular contours, and the integrity of it’s basement membrane was continuous. Immunofluorescence and DAPI staining showed that collagen IV, LN, FN, HSPG2, elastin were maintained during decellularization process. The DNA content in the scaffold was measured in order to demonstrate if there was the cells in the scaffold. The agarose gel electrophoresis showed that DNA bands were less in HTC group than those in control, H,HT groups. Conclusion The study shows that decellularized kidney maintains critical three-dimensional structure and contents by treating with Triton X-100, SDS sequentially at appropriate concentration and ratios. This method is ideal for the experiment of kidney tissue engineering and repair. References | Related Articles | Metrics

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Role of Huangqi combined with Danshen in the denervated skeletal muscle atrophy PEI Yan-hong LIU Kun-xiang* 2014, (2):  273-277.  doi: 10.3969/j.issn.0529-1356.2014.02.025 Abstract ( )   Objective To investigate the therapeutic effect of Huangqi and Danshen in the denervated skeletal muscle atrophy.Methods Eight weeks old male SD rats were randomly divided into the control groups and the treatment groups. To establish the denervated right gastrocnemius atrophy model,about 1cm of the right sciatic nerve was cut off.After operation,1.5ml of a mixture of Huangqi and Danshen was injected into the treatment groups daily via intraperitoneal injection.The control groups were without any treatment.The right gastrocnemius muscle was removed at the second week and the third week after operation.The muscle wet weight,cross section area of myocytes,total protein content,cell apoptosis rate, the mRNA and protein expression levels of FoXO3a, MAFbx were examined. Results Of second week control and treatment groups, the muscle wet weight ratio was 0.62±0.07 and 0.76±0.05; muscle fiber cross-sectional area was 300.24±17.87 and 365.49±24.19; total protein quantity was 63.32±5.30 and 80.28±3.12, apoptosis rate was 20.34± 2.51 and 12.62±1.20, the expressions of FoXO3a protein and mRNA were 0.828±0.090, 0.643±0.090, 24.45±7.21, and 19.38±5.55, the expressions of MAFbx protein and mRNA were 1.224±0.090, 0.956±0.040, 27.45±8.50, and 19.44±5.64. Of third weeks control and treatment groups, the muscle wet weight ratio was 0.35±0.03, 0.59±0.07; muscle fiber cross-sectional area was 253.38±13.32, 314.50±16.74, total protein quantity was 50.34±4.12, 67.70±5.42; apoptosis rate was 33.45±1.71, 17.53±1.26, the expressions of FoXO3a protein and mRNA were 1.963±0.150, 1.236±0.060, 35.62±6.24, and 27.91±7.71. MAFbx protein and mRNA expression were 1.487± 0.050, 1.240±0.123, 43.63±8.22, and 35.39±7.96. Compared between the two groups, the difference was statistically significant (P<0.05). Conclusion Combined application of astragalus Danshen can delay the early rat denervated skeletal muscle atrophy, and its mechanism may be through inhibiting apoptosis, delay the total protein degradation rate, and inhibiting protein expression level and protein degradation pathway of FoXO3a, and MAFbX mRNA. References | Related Articles | Metrics

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Eruption comparison of lower third molars of Chinese recent population DAI Cheng-ping LI Hai-jun* 2014, (2):  283-285.  doi: 10.3969/j.issn.0529-1356.2014.02.026 Abstract ( )   Objective To understand the degeneration of the lower third molars(M3) of Chinese recent population. Methods This study was conducted on 358 human mandibles from China. M3 was divided into 3 types: normal, degeneration and absent. Comparisons were made among different time periods. Results The frequency of the degeneration of the lower third molars was the highest in modern people, and the lowest in Neolithic age.Conclusion The frequency of the degeneration of the lower third molars goes up as time period goes by, which may be caused partly by the obvious size decrease of mandibles. References | Related Articles | Metrics

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Correlation between D6S296, D8S264 gene polymorphism and schizophrenia of Han/ Zhuang people in Guangxi ZHU Sha-sha LI Song-feng* 2014, (2):  286-290.  doi: 10.3969/j.issn.0529-1356.2014.02.027 Abstract ( )   Objective To investigate relationship between D6S296, D8S264 gene polymorphism and schizophrenia of Han, Zhuang people in Guangxi as well as its relevant difference. Methods Han patients (46 cases) with schizophrenia and their immediate family members of the health (49 cases), Zhuang patients with schizophrenia (45 cases) and the health (48 cases) were selected as the research object in Brain Hospital of Guangxi,with healthy families as normal control group. To extract the whole blood DNA and use PCR for DNA amplification, the PCR products with ABI3730XL gene analyzer automatically detect, and statistical genetics method were analyzed. Results D6S296/D8S264 of Han, Zhuang normal control group, each allele frequency distribution were accord with Hardy-Weinberg genetic law of balance (P>0.05). The comparison of Han, Zhuang group of patients with schizophrenia, 264 bp of D6S296 allele frequencies were 2.2% and 18.9% (2/17) respectively, with statistically significant difference (χ 2=13.60, P<0.01);129 bp of D8S264 allele frequency were 18.5% / 8.0% (17/7) respectively, and the difference was statistically significant (χ 2=4.31, P<0.05), and distribution of other allele frequency in each group had no statistical significance (P> 0.05). Conclusion Schizophrenia of Han, Zhuang population in Guangxi may have nothing to do with D6S296, D8S264 polymorphism. References | Related Articles | Metrics

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Pathology and impact of the locus ceruleus in Parkinson’s disease YAO Ning XU Qun-yuan* 2014, (2):  291-296.  doi: 10.3969/j.issn.0529-1356.2014.02.028 Abstract ( )   Objective Parkinson’s disease (PD) is a degenerative disorder of the brain characterized by the degeneration of dopaminergic neurons in the substantia nigra pars compacta (SNc). Recently, however, it has been shown that noradrenergic cells from the locus coeruleus (LC) also degenerate prior to the SNc. Animal studies have disclosed that the electrophysiology and transmitter metabolism of DA neurons as well as its sensitivity to injury factors were abnormal due to LC lesion. The mechanism maybe lies in the function of LC which can accommodate of transmitter in DA neurons, absorb toxic substance and excrete nutrition factors to the SNc area influencing neurons and/or glial cells, leading to promotion of onset of PD. In this article, we reviewed the pathology of the LC in PD patients, the relation between the LC and SNc in laboratory researches on PD animal models, and the protective mechanism of LC to the SNc, intending to help further studies on PD pathogenesis. References | Related Articles | Metrics