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2013, Volume 44 Issue 3
06 June 2013 Previous Issue    Next Issue

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Prenatal alcohol exposure inducing retinal teratogeny and cell apoptosis LI Rui-ling ZHOU Jie WANG Qiang LIU Bin HUANG Xia MA Zhan-you DENG Jin-bo* 2013, 44 (3 ):  297-303.  doi: 0.3969/j.issn.0529-1356.2013.03.001 Abstract ( )   Objective To investigate the effect of teratogeny and cell apoptosis on retina after prenatal alcohol exposure (PAE).Methods A total of 72 mouse models of prenatal alcohol exposure were made. HE staining and immunofluorescent labeling were carried out to visualize the retinal pathology and cell apoptosis. Results High prenatal alcohol exposure increased the rate of retinal malformation; The expression of positive cells in inner granular layer(INL) was costistent with ganglial cell layer(GCL) in the retina with dose-dependency (P<0.05); TUNEL showed in the retina INL and GCL, alcohol induced thecell apoptosis with long-range effects (P<0.05). Conclusion Retinal malformation and cell apoptosis probably are the cause of eye diseases induced by PAE. References | Related Articles | Metrics

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Correlation of draxin expression and neural crest migration in chick embryonic spinal cord at different developmental stage SU Yu-hong* ZHANG San-bing CUI Hui-xian KANG Lin WANG Lei DU Juan YUAN Shi-jun 2013, 44 (3 ):  304-307.  doi: 10.3969/j.issn.0529-1356.2013.03.002 Abstract ( )   Objective To examine the correlation of draxin expression and neural crest migration in chick embryonic spinal cord at different developmental stages.Methods Ten chick embryos at different stages were used. In situ hybridization and immunohistochemistry were used to observe the localization of draxin expression, the migration pathway of neural crest cell-the spatial correlation of draxin expression and the neural crest migration. Living tissue incubation with draxin alkaline phosphatase (draxin-AP) recombinant protein was used to check the binding ability of migrating neural crest with draxin protein. Results With development, the draxin expression and neural crest migration had a very similar anterior-posterior gradient in chick embryonic spinal cord. Draxin was expressed in the dorsal neural tube, roof plate and dorsal tip of dermomyotome which surrounded the migration pathway of neural crest cells. We also found that portion of migrating neural crest cells bound to draxin-AP protein directly. Conclusion Draxin is expressed in the tissues surrounding the neural crest migration pathway and portion of migrating neural crest cells binds to draxin protein directly. We conclude therefore that draxin may be involved in the regulation of neural crest cell migration in chick embryonic spinal cord. References | Related Articles | Metrics

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Expression of brain-derived neurotrophic factor and neurotrophin-3 in post-embryonic development of cerebral cortex of the Chinese alligator ZHENG Lan-rong WU Xiao-bing* WANG Ren-ping ZHOU Yong-kang 2013, 44 (3 ):  308-312.  doi: 10.3969/j.issn.0529-1356.2013.03.003 Abstract ( )   Objective  To investigate sequential changes of brain-derived neurotrophic factor(BDNF) and neurotrophin-3(NT-3) expressed in the post-embryonic development in cerebral cortex of Alligator sinensis. Methods Immunohisto chemistry technique and Western blotting assay were used to detect the temporal expression concerning its variation at location and content of BDNF and NT-3 in the cerebral cortex of the Chinese alligator by postembryonic age of 0 year(newly hatched), 1 year and 2 years. Results Immunohistochemical test showed that BDNF-positive cells were seen at the 4 layers of cerebral cortex from the alligator aged 0 and increased with ages. By 2 years, the cell body for BDNF-positive cells gradually expanded, with the increased nerve processes that were mostly observed at the transition zone of the cell layer and the outer plexiform layer, which was consistent with Western blotting findings. NT3-positive cells were also seen in cerebral cortex of the newly hatched alligator, concentrated in the cell layer and the outer plexiform layer, and gradually increased by 1 year of age, with more intensive positive cells in the outer plexiform layer. By 2 years, NT3-positive cells increased in the molecular layer at lateral zone, and similarly, cell projections tended to increase with ages, which agreed with the results by Western blotting assay. Conclusion Both BDNF and NT-3 are involved in the post-embryonic development in cerebral cortex of Chinese alligator within 2 years of age. References | Related Articles | Metrics

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Effects of acetylpuerarin on the expression of protein kinase C -δ and interleukin-6 in Alzheimer disease rats LIU Min MENG Qing-hui* GUO He WANG Juan-juan 2013, 44 (3 ):  313-317.  doi: 10.3969/j.issn.0529-1356.2013.03.004 Abstract ( )   Objectiv To investigate the effect of acetylpuerarin on the expression of inflammation factors of protein kinase C(PKC-δ) and interferon-6(IL-6) at the hippocampus stereotactic injection of Aβ1-42. Methods Forty Wistar rats were randomly divided into the control group, AD model group,acetylpuerarin low-dose group and high-dose group. AD model rat was produced by bilateral hippocampal injection of Aβ1-42.Learning and memory of the rat was tested through the method of Morris water maze,ELISA was used to determine the expression of IL-6 and Western blotting to detect the changes of PKC-δ. Results Fourteen days after Aβ1-42 injection, Morris water maze escape latency was more extended than pre-surgery and control group, and the times of crossing the platform were significant reduction (P<0.01). After application acetylpueraria for 10 days, in the model group the expression of IL-6 in the serum was significantly higher than the control group,acetylpuerarin low -dose group and high-dose group (P<0.01); The amount of the expression of PKC-δ in the hippocampal homogenate of model group was significantly higher than the control group, acetylpuerarin low -dose group and high-dose group, but the expression of PKC-δ in the acetylpuerarin low -dose group and high-dose group was higher than the control group (P<0.05). Conclusion Acetylpuerarin can significantly improve the learning and memory ability of AD rats. It has obvious preventive and protective effect on Alzheimer disease. The possible mechanism may be that acetylpueariu can regulate and control the Caspase pathway,attenuate the neurotoxicity of β-amyloid peptide, and reduce PKC-δ and IL-6 expression,and thus plays the role of anti-dementia. References | Related Articles | Metrics

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Cell proliferation and synaptophysin expression in the developing mouse lens LI Xue CHEN Wen-jing ZHOU Shu-fang ZHAO Jing-ya DENG Jin-bo* 2013, 44 (3 ):  318-323.  doi: 10.3969/j.issn.0529-1356.2013.03.005 Abstract ( )   Objective To study cell proliferation and the expression of cell cycle protein (cyclin D1) and synaptophysin (SYN) in mouse lens. Methods A total 150 mice were used in the study. HE staining and DAPI staining were performed out to observe the general structure of the mouse lens. Proliferating cell nuclar antigen (PCNA) immunofluorescent staining and 5-bromodeoxyuridine (BrdU) assay were used to mark the proliferative cells in lens. The expression of cyclin D1 protein and synaptophysin in the lens were investigated with cyclin D1 and synaptophysin immunofluorescent staining. Results At about E8, the lens were evolved from the lens placode with single columnar epithelium. Then, the hollow lens bulb developed further and was filled with numerous fibroblasts, which gradually formed lens. BrdU and PCNA-positive cells were mainly distributed in the epithelium of anterior capsule. However, P10 afterward, BrdU positive cells disappeared, while PCNA continued to express in the anterior epithelial cells. Cyclin D1-positive cells were mainly distributed in the epithelium of anterior capsule and the equator of the lens in the embryonic days and early postnatal days. After P5, cyclinD1-positive cells were mainly located in the pre-equatorial area, and only few cyclin D1 positive cells were found in the equator. Synaptophysin protein expressed in the matrix and some cell bodies of equator during the development.Conclusion The cell proliferation plays an important role during the lens development and lens repair. Cell cycle proteins, cyclin D1 is probably involved in regulating the cells’disappear in the equator, therefore the lens’s transparency can be kept. Synaptophysin’s expression in lens has important function in lens’s transparency, through its regulation to calcium flow in cell bodies and matrix of lens. References | Related Articles | Metrics

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Effect of hypoxia-ischemia on the expression of feerroportin 1 in O-2A progenitor cells LIN Qing ZHANG Geng QIU Rong-hui WANG Wei* 2013, 44 (3 ):  324-329.  doi: 10.3969/j.issn.0529-1356.2013.03.006 Abstract ( )   Objective To investigate the FPN1 expression and its role in O-2A progenitor cells after hypoxic-ischemic injury. Methods O-2A progenitor cells were cultured in vitro, indentified with A2B5 antibody and investigated by the localization of FPN1. Hypoxic-ischemic cell models were established by using the oxygen-glucose deprivation (OGD) method and the cell viability was assessed by the CCK-8 method. The expression of FPN1 in O-2A progenitor cells after hypoxia-ischemia was detected by immunofluorescent staining, quantitative real-time polymerase chain reaction analysis and Western blotting analysis. Results FPN1 was localized at the cell membrane, and in the cytoplasm and processes of O-2A progenitor cells. The cell viability decreased with time-dependence after 3hours, 6hours, 12hours and 24hours of OGD (P＜0.05). The FPN1 immunofluorescence intensity of O-2A progenitor cells decreased progressively within 12hours of OGD. The FPN1 mRNA and protein levels downregulated with time-dependence after 3hours, 6hours and 12hours of OGD (P＜0.05). Conclusion The level of FPN1 expression is down-regulated and cell viability decreased significantly with time-dependence after OGD, which suggests that FPN1 may play a role in the hypoxic-ischemic injury of O-2A progenitor cells. References | Related Articles | Metrics

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Inhibition of epigallocatechin-3-gallate on scald-induced macrophage inflammatory protein-2 expression in the skin of mice LI Jia-wen CUI Hai-yan ZHANG Dong-fang XIAO Peng LI Wei-guo* 2013, 44 (3 ):  330-333.  doi: 10.3969/j.issn.0529-1356.2013.03.007 Abstract ( )   Objective To investigate influence of epigallocatechin-3-gallate (EGCG) on expression of macrophage inflammatory protein-2(MIP-2) induced by locally scalded skin in mice. Methods Fity-two Kunming mice were divided into the control, scalded skin, and EGCG groups. Forty-eight mice were used to establish a skin scalded model. The real time quantitative PCR (qRT-PCR) and ELISA were used to evaluate MIP-2 expression in cutaneous tissue at both the mRNA and protein expression levels. Results Both MIP-2 mRNA and protein levels in cutaneous tissue at injury sites increased after skin was scalded, but both decreased after the scalded skin was treated by 0.2g/kg EGCG. Conclusion The data suggest that 0.2g/kg EGCG may suppress MIP-2 expression stimulated by local skin scald, and decrease inflammatory reaction induced by MIP-2 release in cutaneous tissue at injury sites, thus EGCG is able to promote the repair of skin injury elicited by scald. References | Related Articles | Metrics

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Effect of mouse Sertoli cells on human umbilical cord mesenchymal stem cells proliferation ZHANG Fen-xi HONG Yan* LIANG Wen-mei 2013, 44 (3 ):  334-338.  doi: 10.3969/j.issn.0529-1356.2013.03.008 Abstract ( )   Objective To observe the regulatory effect of Sertoli cells (SCs) on proliferation in human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro. Methods hUCMSCs and SCs were isolated, cultured and identified in vitro. A co-culture system was established by culturing SCs in the Transwell insert and hUCMSCs on the plastic plates(the co-culture group). hUCMSCs culture in DMEM-F12 medium was used as the control group. hUCMSCs culture in DMEM-F12 medium supplemented IL-3 and GM-CSF served as the cytokine group. Proliferation, cell cycle analyse and surface marker molecules of hUCMSCs were studied by flow cytometry and immunocytochemistry. Ultrastructures of the cultured cells were observed by transmission electron microscopy. Results Compared with the control, proliferation of hUCMSCs in the co-culture- and cytokine-group increased significantly in the co-culture group (P<0.05). The expression of CD29 and CD105 on the surface of hUCMSCs in the co-culture and the cytokine group also increased. Cell cycle analysis showed that there were a few cells arrested at quiescent phases in the co-culture and cytokine groups, especially in the cytokine group. In the coculture group and the control group，hUCMSCs may displayed the ultrastructural features of the primitive cells by transmission electron microscopy. Conclusion Proliferation of hUCMSCs may be promoted as co-culture with SCs and still kept their stem cell properties after the co-culture. References | Related Articles | Metrics

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Comparison of effect on the induction of three methods of inducing CD73+ adipose-derived mesenchymal stem cells into myocardial cells QI Li-jie GUO Kang LI Qiong GUO Zhi-kun* 2013, 44 (3 ):  339-344.  doi: 10.3969/j.issn.0529-1356.2013.03.009 Abstract ( )   Objective To isolate and purify CD73 positive cell subsets from the mixed adipose-derived mesenchymal stem cells （ADMSCs）, and to compare the differentiation potential of CD73 cell to cardiomyocyte by three methods. Methods ADMSCs of 1-3-month-old mice were isolated and cultured in vitro. ADMSCs subpopulations for CD73+ and CD73- were selected by flow cytometry, and were cultured separately. Two groups of the sorted cells were induced into the myocardium by using 5-aza, myocardial tissue lysate and 5-aza+ myocardial tissue lysate respectively. Immunohistochemistry was used to detect the effect of myocardium after the induction of three ways. Results Unsorted ADMSCs differentiated into adipocytes and ostcoblasts. Sorted CD73+ADMSCs possessed a perfect differentiating potential for cardiaomyocyte. All three induction methods inducted CD73+ ADMSC into cardiomyocytes. The differentiation rate of myocardial cells which induced by 5-aza was (22.99±6.72)%, the rate by myocardium lysate was (14.12±5.42)%, and the rate by 5-aza+myocardium lysate was (26.94±6.11)%. Of the three methods, 5-aza+myocardium lysate was the best method for inducting CD73+ ADMSCs to myocardial cell(P<0.05). Conclusion CD73+ADMSCs may differentiate to cardiomyocyte more easily than CD73-ADMSCs. Chemical induction factors with the simulated myocardial microenvironment in vitro may efficiently induce ADMSCs to differentiate into cardiomyocyte. References | Related Articles | Metrics

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Expression of Nrf2 induced by sodium arsenite in mouse pancreatic β cells YANG Bei* CHEN Xue DING Jin-lan 2013, 44 (3 ):  345-348.  doi: 10.3969/j.issn.0529-1356.2013.03.010 Abstract ( )   Objective To test the expressions of Nrf2 and phase II detoxifying enzyme genes induced by sodium arsenite (NaAsO2) in mouse pancreatic β-cell MIN6.Methods MIN6 cells were exposed to 4μmol/L NaAsO2 for 2, 6, 12, 18, 24hours. Nrf2 protein levels in Nuclear factions, cytosolic factions and whole-cell of MIN6 cells were measured with Western blotting. mRNA expressions of Nrf2 and phase II detoxifying enzyme genes were measured by Real-time PCR. Results Expression of Nrf2 protein was significantly increased after 4μmol/L NaAsO2 exposure did reached the peak at 2hours, then decreased gradually. Nrf2 protein expressions in nuclear factions were consistent with Nrf2 levels of the whole MIN6 cell in the time-dependent fasion. But 4μmol/L NaAsO2 exposure did not induce a robust increase in Nrf2 mRNA. The induction of some phase II detoxification enzyme genes, including NAD(P)H:quinone oxidoreductase 1 (Nqo1), heme oxygenase 1(Homx-1), were time-dependently increased in MIN6 cells. Nqo1 and Homx-1 mRNA expressions were obviously increased after 2hours NaAsO2 exposure and went higher at 6hours, then decreased gradually. Conclusions Upon exposure to NaAsO2, Nrf2 is stabilized and translocated into the nucleus, and then activates transcription of various detoxification enzyme genes in mouse pancreatic β-cells. References | Related Articles | Metrics

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Expression of germ cell nuclear factor and Oct-4 before and after in vitro differentiation of mouse P19 Cell line WANG Ju1, 2 XIONG Cheng-liang GAO Qi-yun LI Hong-gang WANG Miao ZHAO Kai ZHOU Zong-yao 2013, 44 (3 ):  349-352.  doi: 10.3969/j.issn.0529-1356.2013.03.011 Abstract ( )   Objective To detect the expressions of Oct-4 and germ cell nuclear factors(GCNF) before and after inducing P19 cells to differentiation. Methods The P19 cells were induced to differentiation with ATRA treatment.The immunofluorescence staining and RT-PCR techniques were used to detect the expression of GCNF and Oct-4 before and after differentiation of mouse P19. Results The intensities for Oct-4 immunofluorescence positive staining were stronger in non-differentiated P19 cells than that in the differentiated P19 cells. The intensities for GCNF immunofluorescence positive staining were weaker in non-differentiate P19 cells than that in the differentiated P19 cells. Conclusion GCNF is involved in the in vitro differentiation progress of the malignant germ cell tumors. GCNF be inhibited through the Oct-4 gene expression in vitro differentiation of P19 cells. References | Related Articles | Metrics

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Three-dimensional visualization of microvessels related to the peripheral nerve by using microcomputed tomography MAO Yi-hua ZHU Zhao-wei DING Mao-chao TANG Mao-lin* 2013, 44 (3 ):  353-356.  doi: 10.3969/j.issn.0529-1356.2013.03.012 Abstract ( )   Objective To develop a new method for three-dimensional reconstructions of microvessels related to peripheral nerve by using microcomputed tomography(Micro-CT), and to compare the effect of imaging between two different radiopaque materials. Methods Forty SD rats were randomly divided into 2 groups, and then injected with lead oxide-gelatin and microfil respectively. Sciatic nerves were scanned by Micro-CT. The best scanning parameter was set up at 30kVP and 40W. The sectional images were transferred to Dicom format and imported to a personal computer. Three-dimensional reconstructions of microvessels were performed by using Mimics 10.0 software. Results We compared the images of the specimens injected with different radiopaque materials, and found that imaging effect of gelatin-lead oxide was better than microfil. Micro-CT showed that imaging of gelatin-lead oxide was of characteristics such as low cost, low variation and high definition in scanning and reconstruction. Conclusions Microfil has disadvantages such as strict requirements of perfusion pressure, poor imaging quality and other characteristics in microangiography, while the traditional gelatin-lead oxide has its unique advantages in those areas. Micro-CT is a good method in study of microvascular architecture of rat’s peripheral nerve. References | Related Articles | Metrics

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Parietal lobe convexity protuberance in human brain GE Wei-li YU Hong-rong LIAO Zhang-ding HUANG Yi-mei GONG Jian-gu LAN Ling LAO Ming* 2013, 44 (3 ):  357-359.  doi: 10.3969/j.issn.0529-1356.2013.03.013 Abstract ( )   Objective To understand the gyri shape of parietal lobe convexity in human brain. Methods The configuration of gyri on the lateral surface of the parietal lobe of 60 formalinfixed cerebral hemispheres was observed by naked eyes, and measured with vernier caliper.Results There was a protuberance on the convexity of parietal lobe, posterior to lower portion of postcentral gyrus. The area of the protuberance was about 2cm×2cm to 45cm×45cm, 3cm above auricle. The protuberance mainly consisted of the supramarginal gyrus, which was of various shapes arched superoposteriorly, or was a short gyrus dropped superoposteriorly and inferiorly. When the protuberance more obviously protruded,the lower portions of the precentral and postcentral gyri were pushed forwards and their upper portions were pushed backwards thus the whole shape of the precentral gyrus looked and postcentral gyrus looked like “s” shape. Conclusion There is a constant protuberance in the parietal lobe convexity which can be located from the structures inside and outside of skull. References | Related Articles | Metrics

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Comparison between brain magnetic resonance imaging and sectional anatomy of hypothalamic nuclei  ZHANG Rui1 DENG Xue-fei XU Zhao-yang ZHU You-zhi ZHANG Yu MO Zi LUO Xiang-wei HAN Hui* 2013, 44 (3 ):  360-363.  doi: 10.3969/j.issn.0529-1356.2013.03.014 Abstract ( )   Objective To explore the morphology, location and signal of the hypothalamic nuclei in the whole brain magnetic resonance imaging by comparing between magnetic resonance imaging（MRI）and sectional anatomy images of the same specimen of the human adult brain. Methods Six human adult cadaveric heads were examined using MRI and sectional anatomy methods. They were sectioned at horizontal, sagittal and coronal plane，two for each. The collected images from MRI and physical sections of the same brain were prepared. Results In T1 and T2 images, fornix and anterior commissure showed a sharp structure with hypo-signals,of which T1 images were even better. By a landmark such as the fornix or anterior commissure, the paraventricular nucleus in the T1 image showed sharp hypersignals, and slightly sharp hypersignals in T2 images. The mammillary nucleus showed a slightly sharp hypersignals in T1 images, and the mixed signals in the T2 images and the boundary was unclear. The supraoptic nucleus in the T1 image was less clear in boundaries with slightly hypersignals，in T2 images, it was poorly showed. Conclusion Hypothalamic nuclei can be identified in the routine MRI head scan. References | Related Articles | Metrics

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Anatomy of the femoral fascia sheath and its adjacent structures XU Zhao-yang REN Yan-hong TU Li-li XU Sheng-chun* LIANG Liang ZHANG Ming 2013, 44 (3 ):  364-367.  doi: 10.3969/j.issn.0529-1356.2013.03.015 Abstract ( )   Objective  To determine if a femoral nerve “sheath” exists and provide anatomic basis for the lower limb nerve block. Methods Ten sides of lumbar and thigh regions from 5 formalin-fixed cadavers were used in this study. Eight out of 10 specimens were injected with latex using a step-by-step injection method from the distal to the proximal and dissected for topographic anatomy examination. The other two sides from the same body were deep frozen at -35℃ for a week, cut with a bandsaw and used for sectional anatomy examination. Results An intact femoral nerve “fascial sheath” was not observed in the specimens examined in this study. The nerve was surrounded by various fascia-like structures. From the proximal to the distal, those fascia-like structures were the fascia of lumbar plexus, fascial septum of the psoas, the iliac fascia and loose connective tissue covering the femoral nerve. Conclusion At the gross anatomy level, an intact femoral nerve “fascial sheath” may not exist. References | Related Articles | Metrics

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Effects of time and temperature of curing on hardness of organs in silicone plastination ZHENG Wen-xin ZHOU Jia-nan YU Sheng-bo SUI Hong-jin* 2013, 44 (3 ):  368-371.  doi: 10.3969/j.issn.0529-1356.2013.03.016 Abstract ( )   Objective To study the relationships between the hardness of specimen and curing temperature, and time. Methods Thirty specimens of muscle, liver, lung, intestine and brain, 6 each were collected from formalin-fixed human cadavers and processed with the same plastination procedure before curing. During the curing process, the specimens were placed in a sealed chamber, exposed to Hoffen R6 gas and cured for 49 days at different temperature. Results During the curing process, the hardness of specimens increased with time. The liver specimen had medium hardness in 3 days, and the brain in 7 days, and the lung, intestine and muscle in 14 days. The hardness of the liver increased rapidly at 30℃，45℃，60℃ temperature. The hardness of the brain specimen increased rapidly at high temperature. The obvious hardness difference of muscle specimen is not observed in the curing of two weeks among different temperature conditions. After two weeks of hardening, the hardness of muscle increased rapidly at high temperatures. No obvious hardness differences of the specimens of lung and intestine were observed in two weeks during curing at different temperature conditions. Conclusion The low-temperature curing (at room temperature, 30℃) is better for the specimens of liver, intestine, and lung . The specimens of brain and muscle require low temperature curing (at room temperature, 30℃) at first, and then high temperature curing (45℃, 60℃). References | Related Articles | Metrics

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Effect of Chinese traditional medicine on activities of antioxidant enzyme and special protein of kidney in ephedrine addiction mice LI Chong-yang WANG Jin-jin YU Shi-yuan* SUN Jun 2013, 44 (3 ):  372-378.  doi: 10.3969/j.issn.0529-1356.2013.03.017 Abstract ( )   Objective To research the effect of Chinese traditional medicine on ephedrine addiction mice. Methods A model of addiction in ephedrine dependent mice was established by intraperitonealy injected escalation doses ( 2.0, 3.0, 4.0 g/L ) of ephedrine 15 days，and then for withdrawal by irrigate to stomach with escalation doses(20.0，30.0，40.0g/L) astragalus and ligustici chuanxiong Hort decoction and Chinese medicine compound recipe．The samples on withdrawal at 5，10 and 15 days. The content of bloodurea nitrogen(BUN) and malonaldehyde(MDA)，and activities of superoxide dismutase(SOD) and catalase(CAT) were detected by spectrophotometer. The expression of Bax protein and TGF-β1 were examined by immunohistochemical methods. Structures of and the kidney were observed by optical microscopy after irrigation of Chinese medicine. Results SOD and CAT activities the of experimental control group were lower than natural control group. While MDA content increased, the expression intensity of Bax protein and TGF-β1 of the experimental control group were higher than the natural control group. The content of blood serum BUN increased structures of the kidney appeared severity harm. The SOD and CAT activity of kidney in astragalus ligusticum chuanxiong Hort and Chinese medicine compound group increased，while the content of MDA decrease．The degeneration of kidney tissue structure significantly decreased in astragalus ligusticum chuanxiong Hort and Chinese traditional medicine compound groups. The intensity of positive expression of Bax protein and TGF-β1 intensity decreased，The content of blood serum BUN markedly reduced. Conclusion  Astragalus，ligusticum chuanxiong Hort and Chinese medicine compound recipe may improve the ability of cell anti-oxidation, promote the cell repairing and reduce cell damage. References | Related Articles | Metrics

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Development of the retinal vascular system of rodents YAO Huan-ling WANG Tian-shi LIU Ding LI Xiao-fei LIU Jing DENG Jin-bo* 2013, 44 (3 ):  379-385.  doi: 10.3969/j.issn.0529-1356.2013.03.018 Abstract ( )   Objective To study the development of the retinal vascular system in rodents. Methods Immunohistochemistry, gelatin-ink perfusion, and transmission electron microscopy (TEM) were utilized to demonstrate structures of the retinal vasculature and blood-retinal barrier in rodents of different ages（n=12）. Results The rodent retinal vascular system developed from the optic disc after birth and radiated out to cover the entire retina. The superficial vasculature formed on the retinal inner surface, then the vasculature extended into the inner and outer edges of the retinal inner nuclear layer, forming two deep-layer vasculature that was parallel to the superficial vasculature. The blood-retinal barrier was initially composed of naive endothelial cells, a basal membrane of non-uniform thickness, and multiple thick layers of glial cells terminal feet, eventually it developed into a mature blood-retinal barrier composed of a smooth endothelial layer, even basal membrane, and relatively thin terminal feet. Conclusion The maturation of rodent retinal vasculature is spatially and temporally is consistent with the function performance of retinal photosensitivity. The mature blood-retinal barrier endows certain anti-infectious immunity to the retina. References | Related Articles | Metrics

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Nano-hydroxyapatite-collegen-chitosan substance compound with bone marrow mesenchymal stem cells to repair tibia defect in mice LI Yang FANG Yan BAI Shu-ling* TIAN Xiao-hong TONG Hao HOU Wei-jian 2013, 44 (3 ):  386-392.  doi: 10.3969/j.issn.0529-1356.2013.03.019 Abstract ( )   Objective To use nano-hydroxyapatite-collegen-chitosan compounded with bone marrow mesenchymal stem cells (BMSCs) to repair tibia bulk defect in mice, and to investigate the application effectiveness of the composite to the long bone defect. Methods BMSCs of mice were isolated and cultured. Biological characteristics of the stem cells were detected. The nano-hydroxyapatite-collegen-chitosan was prepared. The structure, poriness and degradation rate of the composite were observed under a scanning electronic microscope（SEM）. Male mice with a 5 mm tibia defect were randomly divided into three groups: group A treated with the nano-hydroxyapatite-collegen-chitosan compound with BMSCs; group B with the nano-hydroxyapatite-collegen-chitosan and group C with the nanoscale hydroxyapatite compound with BMSCs. At 8 weeks after restoration, the mice were anesthetize and samples were collected. HE staining and X-ray were used to observe the osteogenesis and healing process in the defect area. Results The BMSCs presented fibroblast morphology and expressed the surface markers of mesenchymal stem cells which were of the biological characteristics of stem cells. The nano-hydroxyapatite-collegen-chitosan had abundant pores and poriness with a relatively even diameter of 100-200μm and an appearance ratio of 72.3%-92.7% (an average of about 82.5%). The degradation rate was（60.3±5.4）% . There were extensive communications inside the scaffold. At the 8th week after transplantation, there was no obvious inflammatory response but mass mature osteoid structure, and the scaffold degraded mostly in group A. X-ray showed that the defect area had new callus，increased new bone density and the bridge grafting in group A. The above featrues were less obvious in groups B and C than that in group A(P＜0.01). Conclusion Nano-hydroxyapatite-collegen-chitosan compounded with BMSCs works well for repairing the tibia bulk defect in mice. References | Related Articles | Metrics

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Expression and function of tyrosine kinase receptor KIT in mouse colonic mucosa epithelium KANG Qian XUE Hong SUN Hai-mei LI Xiao-shuang ZHANG Guo-quan LIANG Yuan-jing* ZHOU De-shan* 2013, 44 (3 ):  393-396.  doi: 10.3969/j.issn.0529-1356.2013.03.020 Abstract ( )   Objective To investigate the expression and function of KIT in mouse colonic mucosa.Methods Western blotting and RT-PCR techniques were used to detect the expression of c-kit gene and KIT in colonic mucosa in C57BL/6 mice (wild-type). Immunofluorescence staining was used to examine the distribution of KIT-positive cells in colonic mucosa epithelium of C57BL/6 mice (wild-type) and N-ethyl-N-nitrosourea (ENU)-induced c-kit point mutant homozygous mice (Wads m/m).Intraperitoneal injection of 5-bromo-2-deoxyuridin (BrdU, 30 mg/kg) was used to observe and compare the BrdU-positive cells in distal colonic mucosa epithelium in wild-type and Wadsm/m mice, and to evaluate the effect of KIT on the renewal of colonic mucosa epithelium. Results The result of RT-PCR indicated that c-kit gene was expressed in colonic mucosa.Western blotting showed that KIT was expressed in colonic mucosa. Immunofluorescence staining showed that KIT-positive cells existed in the epithelium of colonic mucosa and were mainly located in the lower part of crypts. BrdU incorporation indicated that the renewal of the epithelium of colonic mucosa in homozygous mice was significantly slower than that in wild-type mice. Conclusion C-kit gene and KIT protein are wildely expressed in the colonic mucosa epithelium and might be involved in the renewal of the colonic epithelium. References | Related Articles | Metrics

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Establishment of a mouse embryonic germ cell line and preliminary study of the expression of imprinted genes HU Jing ZHAO Qiao-shi GU Yan-li BAI Guang-yu WU Xi LEI Lei* 2013, 44 (3 ):  397-402.  doi: 10.3969/j.issn.0529-1356.2013.03.021 Abstract ( )   Objective  To establish a mouse embryonic germ cell (EGCs) line and to detect the expression of imprinted genes in order to provide basic information for further study and application of embryonic germ cells. Methods EGCs were isolate from primordial germ cells collected from the genital ridge of 12.5 days postcoitum (dpc). The pluripotent characteristics of the established EGCs were detected by alkaline phophatase (AKP) staining, immunofluorescent detection of mouse embryonic stem cells (ESCs) surface antigens, and cell differentiations in vivo. The expressions of several patrilineal and matrilineal imprinted genes, such as Ins2, Lgf2, H19, Lgf2r and so forth, were also detected by quantitative reverse transcriptase-polymerase chain reaction in both EGCs and ESCs. Results The EGCs showed positive for alkaline phosphatase. The pluripotency marker Oct4 and the cell surface marker SSEA-1 were also shown in EGCs cells. Karyotype analysis indicted that EGCs had normal 40 chromosomes, and differentiated into the tissues presenting three germinal layers derivations in vivo, suggesting that embryonic germ cells had pluripotent characteristics. Real-time PCR showed that the expression levels of imprinted genes in EGCs were significantly highter compared with those in ESCs. Conclusion The genomic imprinting memories in EGCs generated from primordial germ cells which collected from the genital ridge of 12.5 dpc are completely erased. References | Related Articles | Metrics

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Expression and significance of autophagyrelated proteins LC-3 and Beclin-1 in silicosis occurrence in rats ZHAO Man-man LI Ran DU Ling-jie WANG Hai-tao TIAN Yan-xia CUI Jian-zhong GAO Jun-ling* 2013, 44 (3 ):  403-408.  doi: 10.3969/j.issn.0529-1356.2013.03.022 Abstract ( )   Objective To investigate the expression of autophagy-related proteins LC-3 and Beclin-1 in the rat silicosis model and its mechanisms, and to study the molecular mechanism of silicosis formation from cells autophagy perspective. Methods Ninety healthy male SD rats were randomly divided into three groups: control group, model group and autophagy inhibitor 3-methyladenine (3-MA) group (n=30). The rat silicosis model was made by one-time infusion of the silica dust suspension method, using the non-exposed tracheal intubation, and was given the 3-MA. The rats were sacrificed on the 1st, 3rd, 7th, 14th and 28th day after the modeling. The morphological changes of lung tissue were observed by HE and Masson staining.The expression of LC-3 and Beclin-1 were detected by means of immunohistochemistry and Western blotting. Results There were alveolitis change, silicosis nodules formation and collagen deposition in the model group.The intensities of immunohistochemical positive reaction for LC-3 and Beclin-1 increased at all time points in the model group (P<0.05). LC-3 and Beclin-1 expressions began to increase at the 1st day, peaked at the 14th day(P＜0.05), and decreased at the 28th day, but higher than basal expression. Compared with the model group, alveolitis change, the area and number of silicotic nodules and collagen deposition decreased, and the expression of LC-3 and Beclin-1 decreased at all time points in the 3-MA group(P＜0.05). No significant changes in the trend of each point in time was observed. Conclusion Autophagy may be involved in the pathological process of silicosis in rats, and played an important role in the occurrence and development of silicosis. References | Related Articles | Metrics

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Parthanatos involved in tubular damages of the kidney in rats with type 2 diabetics LI Shu-ling YAN Wen-zhu* 2013, 44 (3 ):  409-412.  doi: 10.3969/j.issn.0529-1356.2013.03.023 Abstract ( )   Objective To observe the parthanatos involved in tubular cell damages of kidney in rats with type 2 diabetics. Methods Animal models of type 2 diabetics were established. Light microscopy, electron microscopy, immunohistochemistry and Western blotting were used in this study. Results A large number of glycogen accumulation, cell swelling, and partial necrosis, mainly for the vacuolar necrosis were seen in early type 2 diabetes renal tubular cells. Immunohistochemical staining of PARP-1 showed that the positive cells were found in epithelium of renal tubules of type 2 diabetic kidney. Compared to the control kidney, PARP-1 expressed much stronger by immunoblot analysis. Conclusion Necrosis and parthanatos are involved in renal tubule damages induced by type 2 diabetes in rats. References | Related Articles | Metrics

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Six behavioral traits of lateral functional dominance in Han nationality of Liaoning province LI Wen-hui XI Huan-jiu* Lü Po ZHAO Hong 2013, 44 (3 ):  413-418.  doi: 10.3969/j.issn.0529-1356.2013.03.024 Abstract ( )   Objective To investigate the distribution frequency of 6 behavioral traits of lateral functional dominance in Han nationality of Liaoning, and to collect data for the Chinese physique survey. Methods Six behavioral traits of lateral functional dominance including hand clasping, handedness, arm folding, leg folding, foot preference and stride type of the Han nationality in Jinzhou of Liaoning province were analyzed. Results The hand clasping, handedness, arm folding, leg folding, foot preference, stride type of Rtype rate were 52.37%, 91.32%, 45.53%, 68.16%, 92.89% and 51.32%, respectively. No sexual difference was oberved. Except arm folding, the percentage of R-type on other 5 traits was much higher than that of L-type. Han nationality in Liaoning had more than four traits which appeared significant differences from that of compared with Bouyei, Dong and Miao. The results of cluster analysis showed that 6 traits of Han ethnic in Liaoning were similar with those of Mongoloid, Oroqen and Daur. There was the higher correlation among 6 traits. In each pair of the traits, the percentage of RR type was much higher than that of LL type. Conclusion The R-type rates of hand clasping, handedness, foot preference, stride type in Liaoning Han ethnic are in the medium level, and that of arm folding and leg folding are at a lower level, compared with other ethnic groups from China and overseas. References | Related Articles | Metrics

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Association of polymorphisms of rs479200 and rs480902 in EGLN1 gene with hypoxia adaptation in high altitude in Tibetans Lü Po FAN Jie XI Huan-Jiu* 2013, 44 (3 ):  419-422.  doi: 10.3969/j.issn.0529-1356.2013.03.025 Abstract ( )   Objective To investigate the association of single nucleotide polymorphisms(SNPs) of rs479200(C/T) and rs480902 (T/C)—EGLN1 with hypoxia adaptation in high altitude in Tibet. Methods The blood samples were chosen from 150 Tibetans in Tibet high altitude region and 150 Han nationality people in Liaoning province, and genomic DNA was extracted from the blood samples. The single nucleotide polymorphisms of EGLN1 gene were examined by restriction fragment length polymorphism-polymerase chain reaction(RFLP-PCR). Results The allele(C) frequency of EGLN1 gene rs479200 in Tibetans and in Han nationality was 71.33% and 38.17%,and the allele(T) frequency of rs480902 was 66.67% and 36.67%. The genotype frequency of EGLN1 gene rs479200 in Tibetans and in Han nationality was 6.67% and 56.67% in TT genotype,29.33% and 33.33% in TC genotype and 64% and 10% in CC genotype. The genotype frequency of rs480902 was 60.67% and 9.33% in TT genotype,30.66% and 28.67% in TC genotype and 8.67% and 62% in CC genotype. TT genotype frequency of rs479200 in Tibetans was lower than that in Han nationality (P<0.01),while CC genotype frequency of rs479200 in Tibetans was higher than that in Han nationality(P<0.01). No significant difference in CT genotype frequency of rs479200 and rs480902 was observed between two groups respectively. Conclusion Polymorphisms of EGLN1 gene rs479200 and rs480902 are associated with hypoxia adaptation in high altitude in Tibetans．CC genotype of rs479200 and TT genotype of rs480902 may be benefit to hypoxia adaptation. References | Related Articles | Metrics

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Somatotype of hoopster in sport school in Panjin of Liaoning province GU Ping XI Huan-jiu* LI Wen-hui 2013, 44 (3 ):  423-426.  doi: 10.3969/j.issn.0529-1356.2013.03.026 Abstract ( )   Objective To study somatotype of hoopsters in the sport school in Panjin, Liaoning, in order to provide a simple effective basis for basketball player selection. Methods A total of 98 hoopsters aged 16 to 20 years old were randomly selected from in the sport school in Panjin of Liaoning, with the informed consent. The control group was the middle school students with the same age. Ten indexes such as height, weight, etc were measured and evaluated by the Heath-Carter anthropometric method. Results The average somatotype of male hoopster was 3.10-4.52-2.77, the female mesomorphic endomorph was 4.07-3.82-2.77. Male and female hoopster height/weight1/3 (HWR) (mean 43.83,42.71) and male the female ordinary high school students (mean to 42.43,41.23) compared with a significant difference (P<0.05). Average body mass index(BMI) of the male and female hoopster were 23.30,22.45 ordinary high school students (male and female respectively 22.12,20.57) there was a significant difference (P<0.05). Conclusion Somatotype of male hoopster is of balanced mesomorph, the female is of mesomorphic endomorph. Compared with middle school students, somatotype of hoopster has a concentrated distribution relatively. References | Related Articles | Metrics

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Effects of different freezing methods on the follicular viability of large pieces of human ovarian tissues ZHAO Shuo ZHAO Shu-qin SHI Qing QIN Xiao-min HAN Yi-long CHAO Lan DENG Xiao-hui* 2013, 44 (3 ):  427-432.  doi: 10.3969/j.issn.0529-1356.2013.03.027 Abstract ( )   Objective To study effects of cryopreservation of large pieces of human ovarian tissues and assess the follicular viability after vitrification and two-step freezing. Methods Ovarian tissues from 15 patients were collected. The size of each tissue block was about 15 mm×15 mm×2mm. The tissue block was cut into three pieces and randomly grouped as fresh(A), vitrification (B), and two-step freezing (C) groups. All the pieces were cultured in vitro after thawing. The viability and proliferative capacity of tissues were evaluated by in vitro production of estrogen and progestogen, histological analysis and immunohistochemical staining of nuclear proliferation-associated antigens, Ki67 and B cell lymphoma/lewkmia-2（Bcl-2）. Results The ratios of normal follicles were the highest in group A(91.9%), median in group C (82.9%) and the lowest in group B(71.3%) (P<0.05). The ratios of morphologically normal primordial and primary follicles in group C were 86.8% and 54.4% which were higher than 73.8% and 44.4% in group B(P<0.05), respectively. Difference of hormonal activity was not found among three groups（P>0.05). Positive expression of Ki67 of group C（73%）was higher than group B（50%）and group A（55%）(P<0.05), where as that of group A was lower than group B (P<0.05).There was no difference of positive expression of Bcl-2 staining between group A（57%）and group C（61%）(P>0.05), but the ratio of positive Bcl-2 expression of group B（42%）was lower than groups A and C (P<0.05). Conclusions The frozen/thawed large pieces of human ovarian tissues can reserve their vitality and the tissue with two-step freezing is better preserved than the tissues with vitrification. References | Related Articles | Metrics

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Two techniques to locate the dominant perforator vessels DING Mao-chao* ZHAO Li-na MAO Yi-hua CUI Huai-rui CHEN Shi-xin MEI Jin TANG Mao-lin 2013, 44 (3 ):  433-436.  doi: 10.3969/j.issn.0529-1356.2013.03.028 Abstract ( )   Objective To provide morphology data that enable to locate perforator vessels and flap design. Methods Two fresh unfixed corpses underwent whole body bismuth oxide and polyvinyl alcohol injection, and then scanned by spiral CT. The CT data was imported into the Mimics imagine workstation to locate the dominant perforator and flap design by volume rendering and dynamic reconstruction. Results Two methods were able to locate the perforator. Rapid direct volume rendering on perforator vessel locating was rapider, but it was not good to show adjacent structures. The dynamic reconstruction method was used for perforator locating and flap design, and also showed adjacent layers clearly with threshold adjustments. Conclusion Rapid direct volume rendering is a quick and simple way to show the locating of the perforating branches of the source artery in various parts. Dynamic reconstruction method is able to display the adjacent structures with multi colored forms.The best way to locating perforators is combining these two methods. The results provide the morphological basis for designing and simulating the flap. References | Related Articles | Metrics

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Therapeutic research on triple negative breast cancer stem cell mammospheres ZHAO Le LIANG Chen LI Qian PEI Xiao-hua* 2013, 44 (3 ):  437-440.  doi: 10.3969/j.issn.0529-1356.2013.03.029 Abstract ( )   Triple negative breast cancer（TNBC）is characterized by estrogen receptor(ER) negative, progesteronereceptor(PR) negative, and human epidermal growth factorreceptor-2(HER-2) negative histochemical staining. TNBC has the features of high malignant degree, strong invasion, prone to distant metastasis and poor prognosis. The endocrine therapy and targeted therapy are generally not satisfied. Finding a effective treatment is a key point in clinical research. Recent studies have shown that occur rence and development progress of TNBC may be related to breast cancer stem cells. Mammosphere culture is a very good method of collection of breast cancer stem cells, and is used very widely in stem cells of TNBC. References | Related Articles | Metrics