目的 探讨正常和热应激条件下，葡萄糖转运蛋白1（GLUT1）和葡萄糖转运蛋白2（GLUT2）在成年猪睾丸的表达和定位。 方法 性成熟长白公猪9头，随机分为3组。局部阴囊热刺激组（n=3），用自制电热毯置阴囊42 ℃加热1 h；环境热应激组（n=3），每天置于37～40 ℃猪舍环境3 h，连续7 d，每天于热处理结束后，将实验猪驱赶回21～25 ℃猪舍环境；对照组（n=3），饲养在21～25 ℃猪舍环境。局部热刺激6 h后和环境热应激处理结束24 h后，手术摘除双侧睾丸。用Real-time PCR、Western blotting和免疫组织化学技术检测猪睾丸组织内GLUT1和GLUT2的表达。 结果 Real-time PCR和Western blotting结果显示，与对照组相比，环境热应激组GLUT1蛋白和mRNA的表达差异不显著，局部阴囊热刺激组GLUT1蛋白和mRNA表达显著升高；环境热应激组和局部阴囊热刺激组，GLUT2蛋白和mRNA表达均显著升高。免疫组织化学结果发现，热处理前后，GLUT1蛋白在曲精小管内定位于精母细胞和圆形精子细胞；环境热应激组GLUT1蛋白染色与对照组相比，无明显差异，局部阴囊热刺激后，GLUT1染色变深，表达升高。热处理前后，GLUT2蛋白在曲精小管定位于生精细胞和支持细胞，环境热应激和局部阴囊热刺激导致GLUT2染色变深，表达升高。 结论 葡萄糖转运蛋白GLUT1和GLUT2表达于猪睾丸曲精小管，环境高温和阴囊局部热刺激导致GLUT1和GLUT2在猪睾丸的表达水平改变，提示这两种葡萄糖转运蛋白在猪精子发生过程中发挥重要作用。

目的 探讨白藜芦醇对NIH3T3细胞Shh信号通路的影响。 方法 实验分为对照组和白藜芦醇组。白藜芦醇处理NIH3T3细胞24 h。CCK-8法测定细胞活力，免疫荧光法检测初级纤毛蛋白（Ac-tu）、Smo和Gli-1的表达，Western blotting法检测Shh、Ptc、Smo、Gli-1蛋白的表达。 结果 1. 0.5和1 μmol/L白藜芦醇组（0.679±0.047, 0.774±0.054）细胞活力分别较对照组（0.585±0.039）明显增强（P＜0.05），其中以1 μmol/L白藜芦醇组最强。之后随白藜芦醇浓度（10、20、40、80 μmol/L）的增高，细胞活力逐渐降低（0.428±0.043, 0.395±0.031, 0.373±0.017, 0.361±0.016），且均较对照组明显减弱（P＜0.05），而0.1 μmol/L（0.602±0.065）和5 μmol/L组（0.556±0.041）细胞活力与对照组比较差异无显著性（P>0.05）。2. NIH3T3细胞表达Ac-tu、Shh、Ptc-1、Smo、Gli-1从蛋白，有1根初级纤毛，Smo和Gli-1蛋白位于细胞质。白藜芦醇处理24h时，Gli-1从细胞质进入细胞核，Smo从细胞质进入初级纤毛，且Shh（0.756±0.659比0.441±0.769，P＜0.05）、Ptc-1（0.655±0.347比0.351±0.026，P＜0.05）、Smo（0.779±0.064比0.451±0.035，P＜0.05）和Gli-1（0.856±0.044比0.560±0.058，P＜0.05）蛋白表达较对照组高。 结论 白藜芦醇可能通过激活Shh信号通路增强NIH3T3细胞的活力。

目的 探讨索拉非尼对大鼠佐剂性关节炎(AA)的抑制作用。 方法 36只雄性SD大鼠均分为6组，除正常组外，其余各组大鼠均制备AA模型。足容积法检测AA大鼠继发侧足爪容积；流式细胞术检测外周血CD4⁺及CD8⁺T细胞亚群的变化；免疫组织化学链霉卵白素-过氧化物酶法(SP)检测滑膜组织微血管密度(MVD)的改变。 结果 与模型组相比，索拉非尼组大鼠足爪容积下降，滑膜组织MVD减小，其中索拉非尼(20、40mg/kg)组可使外周血CD4⁺T细胞比例降低，CD8⁺T细胞比例增加，差异均有统计学意义(P<0.05)。 结论 索拉非尼具有抑制大鼠AA效应，该作用可能与索拉非尼引起AA大鼠外周血CD4⁺, CD8⁺T细胞亚群的偏移以及降低滑膜组织MVD有关。

巨噬细胞在体内分布极广，具有高度可塑性，并在机体的发育及内环境的平衡中起到重要作用。应激条件下，巨噬细胞发生极化，极化后的巨噬细胞亚型可以细致调节及回应各种不同的刺激，对炎症反应或疾病的发展及组织器官的修复程度起着关键性的作用，具有重要的临床应用价值。我们在文中概述了巨噬细胞的极化分类，作用机制及不同环境下巨噬细胞的功能转换，为巨噬细胞极化研究提供理论基础。

目的 分析合并颈椎生理曲度改变的中青年眩晕患者的相关危险因素。 方法 收集2016年6月～2017年3月在新乡医学院第三附属医院神经内科住院的中青年眩晕患者病历，共78例，其中男性32例，女性46例。根据是否有颈椎生理曲度改变分为颈椎生理曲度正常组（n=22）和颈椎生理曲度异常组（n=56），比较两组的既往病史、实验室检查，对结果进行统计学分析。 结果 空腹血糖（FBG）、糖化血红蛋白（HbA1c）、同型半胱氨酸（HCY）、吸烟史及职业组间差异有显著性（P＜0.05）；性别、年龄在两组之间差异无显著性（P＞0.05）。 结论 合并颈椎生理曲度改变的中青年眩晕患者中FBG、HbA1c、HCY、吸烟史、糖尿病及职业是常见的危险因素。

Objective To approach the changes of advanced glycosylated endproducts (AGEs), transforming growth factor(TGF-β1) and connective tissue growth factors(CTGF) in the lungs of experimental diabetic rats and their relationship. Methods Totally 48 male SD rats were divided into the diabetes mellitus（DM） group and the control group，with 24 rats in each group.The DM rat model was made by the injection of streptozocin (60mg/kg) into the caudal vein. The rats were killed and the lungs were taken at the end of the 4 th, 12 th and 20th weeks respectively after the models were established. The changes of AGEs,CTGF,TGF-β1 in rat lungs were observed with immunohistochemical assay and the image was analyzed. Results A great quantity of AGEs positive cells were observed in the alveolar epithelial cells, bronchial mucosal epithelium, angio-endothelial cells and smooth muscle cells of the DM rats. The average gray(AG) was lower than that of the controls（EM>P/EM><0.05） and decreased with the DM course（EM>P/EM><0.01）. In the 4_week DM rats, there were a few CTGF and TGF-β1 positive cells in the bronchial mucosal epithelium, angio-endotheliial cells and lung interstitial cells. In the 12_and 20_week DM rats, there were a great many CTGF and TGF-β1 positive cells. The AG was lower than that of the controls（EM>P/EM

Objective To investigate the effects of transplantation of frozenthawed ovarian tissues on the maturation, fertilization and further developmental potency of oocytes retrieved from grafts. Methods Twenty five New Zealand white female rabbits were divided into three groups randomly, group 1(EM>n/EM>=5), control group; group 2 (EM>n/EM>=10), fresh ovarian tissues were autologously transplanted into the mesometrium; and group 3(EM>n/EM>=10), frozenthawed ovarian tissues were autologously transplanted into the mesometrium. Three months after the transplantation, rabbits were stimulated with folliclestimulating hormone and oocytes were retrieved from antral follicles 13 hours after human chorionic gonadotropin injection. In vitro maturation (IVM) and intracytoplasmic sperm injection (ICSI) were performed to evaluate the fertility potency of the oocytes from frozen ovarian grafts. Results The number of retrieved oocytes in group 2 and 3 were lower than those of the control group (P<0.05); But no significant differences were observed between group 2 and group 3 (EM>P/EM>>0.05); There were no significant differences both in the percentage of immature oocytes and the maturation rate after IVM, among the 3 groups (EM>P/EM>>0.05); Also, among the 3 groups or in each group, the fertilization rate, cleavage rate and blastocyst formation rate showed no difference, no matter the oocytes matured EM>in vivo/EM> or EM>in vitro/EM> (EM>P/EM>>0.05); The blastocyst formation rate derived from oocytes that matured in vitro was significantly lower than oocytes that matured EM>in vivo/EM> (EM>P/EM><0.05). Conclusion

Objective To evaluate the CT features of the coronary artery in normal adults and to measure the diameter of coronary artery using 16-slice computed tomography. Methods 16-slice CT coronary angiography was performed in this study. Totally 104 cases whose coronary arteries were normal were divided into 3 groups according to age and dominant pattern of coronary artery. The diameter of right artery (RCA), including the proximal, middle and distant segments were measured by using CT, so was the diameter of left main artery (LM), left anterior descending artery (LAD) including the proximal, middle and distant segments, and left circumflex artery proximal and distant segments. Results The diameters of coronary arteries increased with the age. There was a difference of LM diameter between the elder group and the youth group and middle group. However there was no difference in the youth group and the elder group. No difference were detected from other coronary artery segment’s diameter. The diameter of LM in left dominant pattern was the largest, and it showed a difference when compared with the other two groups, bt there was no difference between the latter two groups. The diameters of LAD and LCX in left dominant pattern were the largest, and that of in right dominant pattern were the smallest. The diameter of LAD had no difference among the three groups. The diameter of LCX proximal segment had no difference between the balanced dominant pattern and right dominant pattern, but there was a difference compare to the left dominant pattern. The diameters of LCX distant segment showed differences between any two groups. The diameter of RCA in the right dominant pattern was the largest. The diameters of RCA proximal segments showed no difference between the balanced dominant pattern and left dominant pattern, and there was difference among any other groups.Conclusion MSCT can display coronary artery from any angle and position. The diameter of coronary artery segment increases with the increase of age. Coronary artery size measured by MSCT can provide valuable information for coronary disease.

Objective To study the distribution of CD2-associated protein (CD2AP) in normal renal cell lines and its interaction with nephrin and F-actin in podocytes. Methods The human mesangial cells (HMC) and HK-2 were cultured in DMEM. Conditionally immortalized murine podocyte cell line was cultured in RPMI 1640 The expressions of CD2AP and nephrin in podocytes were examined by RT-PCR and Western blotting. The distribution of CD2AP in HMC, HK-2, differentiated and undifferentiated podocytes was observed by laser scanning confocal microscopy. The coexistence of CD2AP with nephrin and F-actin in undifferentiated podocytes were also detected. Results CD2AP was distributed within the cytoplasm and perinulcear region of HK-2 and undifferentiated podocytes, but was absent in HMC cells. Its distribution profile changed and presented as peripheral accumulation when podocytes were put into differentiationpermissive conditions. CD2AP was located together with nephrin and F-actin in podocytes. Conclusion CD2AP can be detected in epithelial-originated renal cells. The alteration of distribution profile of CD2AP indicates it may participate in the process of podocytes differentiation and be involved in the regulation of slit diaphragm and cytoskeleton.

Objective To further explore the mechanism by which bone morphogenetic protein 4（BMP-4）might be involved in hematopoietic differentiation of the yolk sac. We observed the expression of BMP-4,CD34, CD133 and tyrosine kinase receptors（KDR） in the blood island of the yolk sac at embryonic 3 to 8 weeks. Methods Gene expression was analyzed by RT-PCR and the presence of BMP-4, CD34, CD133 and KDR proteins was confirmed by immunohistochemistry in 57 human embryos. Results In the human yolk sac, we found that BMP-4 was expressed at high levels from the 16th day to the 7th week, and decreased quickly after week 7 The results showed that KDR, CD133 and CD34 largely appeared on the 21st and 30th day, then increased at the 6th week, and decreased quickly after week 7. Furthermore, Ihh，SCl, GATA-1, GATA-2 and PU.1 mRNAs showed that PU.1 was not expressed on the 16th day; however, other factors were expressed all the time. Conclusion The distribution of BMP-4,KDR,CD34,CD133 and transcription factors expression highly suggested that BMP-4 was secreted from the yolk sac which might exert its effects on the specification of human hemangioblast and hematopoietic ste

Objective To investigate whether human placenta tissue (PT) CD133SUP>+/SUP> cells possess the high proliferative potential colony forming cells(HPP-CFC) and analyze its properties in order to verify that human PT contains primitive hematopoietic stem/progenitor cells(HSPC). Methods Single cell suspension liquid of human PT was prepared by mechanical method. After that the mononucler cells(MNC) from PT was separated by Histopaque-1007 agent. CD133SUP>+/SUP> cells contained in MNC were isolated and purified by magnetic activated cell sorting(MACS) method. After 28 days of HPP-CFC expansion culture of CD133SUP>+/SUP> cells, the frequency and morphology of HPP-CFC were counted and observed. Phenotypes of purified CD133SUP>+/SUP> cells and HPP-CFC were analyzed by flow cytometer(FCM). In parallel, human umbilical cord blood(UCB) samples underwent the same protocols for comparison. Results After 28 days of culture the expansion fold of PTSUP>-/SUP>CD133SUP>+/SUP> cells was 266, lower than that of UCB-CD133SUP>+/SUP> cells which was 362 (P<0.01). The number of HPP-CFC in CD133SUP>+/SUP> cells derived from human PT and UCB were (32.4±11.2)/5×10SUP>3/SUP> and (17.7±5.7)/5×10SUP>3/SUP> respectively, the number of the former obviously higher than that of the latter (P<0.01). FCM analysis results showed that, except for CD133SUP>+/SUP>CD34SUP>-/SUP> subset from UCB-CD133SUP>+/SUP> cells, the proportion of CD133SUP>+/SUP>CD34+, CD133SUP>-/SUP>CD34SUP>+/SUP> subsets both from PT- and UCB-CD133SUP>+ /SUP>cells and CD133SUP>+/SUP>CD34SUP>- /SUP>subset from PT-CD133SUP>+/SUP> cells were decreased through 4 weeks of culture BR>.Conclusion Human PT-CD133SUP>+ /SUP>cells can form HPP-

P>Objective To investigate the distribution patterns of 15 short tandem repeats (STR) loci (TPOX，TH01，CSF1PO，D19S433，vWA，D18S51，D5S818，FGA，D8S1179，D21S11，D7S820，D3S1358，D13S317，D16S539，D2S1338 ) in Miao ethnic group of Rongshui County in Guangxi Province. Methods The sodium citratedblood specimens were collected from 208 healthy unrelated Miao individuals (man: 102, female: 106) in Rongshui County in Guangxi Province, and then the DNAs from the samples were extracted by phynolchloroform technique. The DNAs were amplified by using AmpFlSTR IdentifilerTM PCR Amplification Kit, and finally the data were detected with 3100 Genetic Analyzer. Result Altogether 5.20 alleles and 11.62 genotypes of 15 STR were found in 208 healthy unrelated Miao individuals of Rongshui County in Guangxi Province. The allele frequency and genotype frequency were 0.004 8-0.466 3 and 0.004 8-0.317 3 respectively. Total discrimination power wa

Objective To investigate the biocompatibility of bone marrow stromal cells (BMSCs) of mice EM>in vitro/EM> with silk fibroin materials and to explore a novel scaffold material to fabricate tissue-engineered nerve with introduction of BMSCs. Methods BMSCs were typically isolated from other cells by adherence to plastic. The mice-derived bone marrow stromal cells were cultured on the substrate of silk fibroin fibers and the cell attachment and growth during culture was observed by using light and electron microscopy. Mice-derived BMSCs were also cultured in the silk fibroin extract fluid. The cell ultrastructure was observed by transmission electron microscopy. MTT test was used to detect cell viability in different media for 12, 24, 48, 72 hours and 7 days respectively (the test was repeated 12 times for each group). Flow cytometry was employed to detect BMSCs cell cycle and phenotypes (the test was repeated 3 times). Results BMSCs cells were tightly attached to the silk fibroin fibers and grew along the silk fibroin fibers; some of them enwrapped the silk fibroin fibers and they exhibited either a spherical or spindle shape. The results of transmission electron microscopy, MTT test and flow cytometry analysis showed that there was no significant difference between BMSCs cultured in the silk fibroin extract fluid and those in plain IMDM medium in their morphology, cell viability, proliferation and phenotypes.Conclusion These data indicate that silk fibroin has good biocompatibility with BMSCs and is also beneficial to the survival of BMSCs without exerting any significant cytotoxic effects on their phenotype; thus it’s a potential scaffold material to fabricate tissue-engineered nerve with introductio

Objective To detect the cytokines in the bone marrow stromal cells (BMSCs) conditioned medium (neurobasal conditioned medium ,N-CM) that can regulate the differentiation of neural stem cells (NSCs) into high proportional neurons and explore the mechanism that BMSCs regulate the differentiation of NSCs.Methods The collected N-CM was divided into two parts(>5kD and <5kD)by means of ultrafiltration after misce bene. The two parts were used to culture NSCs separately, and the effective part that could regulate the differentiation of NSCs was detected by protein microarray analysis. Results The N-CM>5kD could promote the NSCs to differentiate into high proportional neurons, so this part was detected by protein microarray analysis, 7 cytokines CINC-3, CNTF, IFN-γ, IL-1α, MCP-1, TIMP-1and VEGF were detected to up-regulate 1.5 times compared with the neurobasal medium (molecular weight above 5kD). Conclusion The dissolvable molecules excreted by BMSCs could promote the NSCs to differentiate into neurons. Some cytokines with the molecular weight of above 5kD play a crucial role during this process.

Objective To construct five shRNA-expression plasmids and to investigate the expression of EM>Smad2/EM> in TGFβ/ Smads signal transduction treated with shRNA-expression plasmid. Methods Five shRNA-EM>Smad2/EM> DNA sequences from mRNA sequence of mouse EM>Smad2/EM> gene were designed and synthesized. DNA oligonucleotides encoding an appropriate shRNA were inserted to shRNA expression vector respectively. Five shRNAEM>Smad2/EM> expression plasmids were obtained and then transfected into NIH/3T3 cells. The suppressed expression of EM>Smad2/EM> was assessed by RT-PCR and Westernblotting. Results The shRNA-expression plasmid numbered 2.4 could markedly reduce the expression of EM>Smad2/EM>. The suppression effect of the RNAipool composed of four different plasmids was more obvious than that of any single. Conclusion The shRNAexpression plasmids were successfully constructed, which could specifically and effectively suppress the expression of EM>Smad2

Objective Neural stem cells have become a major concern in the current research of neuroscience, for they are involved in the neural injuries and recoveries, the origin of neural tumors, as well as other fields. The study on their ultrastructures, which is still limited at present, is indispensible. To offer more information is the aim of this paper. Methods Neural stem cells/progenitors from human fetal brain tissue were cultivated EM>in vitro/EM> and observed under a scanning electron microscope (SEM) and a transmission electron microscope (TEM). Results Neural stem cells/progenitors presented to be neurospherelike after days of culture EM>in vitro/EM>. The neruospheres were made up of neural stem cells/progenitors and nonfixiform material inside, and cells in neruospheres could be divided into lucent and dark ones according to electron densities. Between adjacent cells as well as on the cytoplasmic sides of the apposed plasma membranes, there were vesiclelike structures. Cell membrane fusions were also observed between some adjacent cells. Single neural stem cell /progenitor was spherical with rough surface under SEM. Many kinds of organellas, e.g. Golgi’s complex and endocytoplasmic reticulum, were underdeveloped in neural stem cells/progenitors, which generally had big, nuclei and scanty cytoplasm. The numbers, types and maturities of cellular organs in different cells were not always identical, which showed their heterogeneities. For instance, both neurofilaments and microtubules could only be observed in a few neural stem cells/progenitors； lysosomes were very abundant in some, but even hardly founded in others. What’s more, autophagosomes at different stages and in differernt formations could be seen in most cells. The nuclei, frenquently containing huge amounts of euchromatin and a small quantity of heterochromatin, mostly were globular, sometimes reniform or lobulated; most neural stem cells/progenitors had only one chromatospherite, seldom two or more, and sometimes no obvious chromatospherite could be seen. Conclusion Developed autophagosomes, vesiclelike structures between adjacent cells as well as on the cytoplasmic sides of the apposed plasma membranes and cellular membrane fusions could be seen in the human embryooriginated neural stem cells/progenitors and

Objective To reveal the distribution and effects of peripheral nerve axolemma ion channels in experimental allergic neuritis (EAN). Methods The alteration of Na+ and K+ channels on peripheral nerves (PNs) in the course of EAN was observed and the relationship between the channels and nerve conduction properties was analyzed by assessing histology and electrophysiology of PNs as well as clinical severity. Results Na+ and K+ channels obviously decreased on day 9 post immunization (p.i.), a time point of disease onset, and quickly became undetectable in next two weeks. Undergoing a slow and incomplete regeneration, neither of the channels regained the normal appearance on day 85 p.i. even if the clinical symptom disappeared several weeks before. Na+ and K+ channels had a synchronous development during disease course and remained a close correlation with the alteration of paranodal myelin. Electrophysiological abnormality kept consistent with the disturbance of PNs just in the acute period of EAN (923d p.i.) and the compound muscle action potential (CMAP) amplitude partly reflected the distribution of axolemma ion channels. Conclusion Loss of axolemma ion channels of PNs might be one of the reasons directly leading to the early symptoms of EAN. As a structural component, the channels were liable to damage and difficult to restore. The clustering and maint

目的 探讨以纤维支气管镜预观察右上叶支气管开口解剖方位，对插右双腔支气管导管（R-DLT）对位的临床意义。 方法 选择插R-DLT成年患者160例，随机分为实验组和对照组，每组80例。实验组病例麻醉诱导后预先行支气管镜检查，主要测量右主支气管长度及右上叶支气管开口在右主支气管横切面的方位(以患者正前方12点钟位置为0度起点,按顺时针增大)。两组按常规方法将R-DLT插入右侧支气管，之后以纤维支气管镜检查调整导管位置。实验组按之前测定的支气管解剖调整导管深度并作适当的旋转，对照组只调整导管深度使蓝色的支气管套囊上缘在隆突之下见到。然后纤维支气管镜改从右管腔插入通过导管的侧孔查看右上叶支气管开口的对位情况，没有进一步调整就能够看到右上叶支气管开口即为初步对位成功。最后适当微调导管，直至能看清右上肺尖段、后段及前段3个开口。比较两组初步对位成功率以及插管失败率。 结果 实验组右主支气管长度(2.29±0.58)cm，其中短于1cm的有2例，占2.5%；右上叶支气管开口在右主支气管横切面方位(94.5±8.3)°，其中有4例（5.0%）明显偏前或偏后。实验组右上叶支气管开口初步对位成功实验组有77例（96.3%），而对照组为62例（77.5%），组间差异显著（P<0.05）。两组各有1例插管失败，占1.25%，均为右上叶支气管开口与隆突距离较近（<1cm）。 结论 预先以纤维支气管镜查看右支气管解剖有助于提高插R-DLT初步对位的准确性，并利于插管前发现右上叶支气管开口变异而选择合适的导管具有重要意义。

P>Objective To establish an immortalized rat astrocyte strain (IAST) expressing enkephalin regulated by doxycycline (Dox) and observe its analysesic effect on rat chronic neuropathic pain. Methods Retrovirus infection method was employed to develop an immortalized rat astrocyte strain expression enkephalin regulated by doxycycline. hPPE gene expression level of IAST/TetOn/hPPE strain was detected by Real time PCR and radioimmunoas say. Its analgesic potential was investigated by mechanical paw withdrawal thresholds after these cells were implanted into the subarachnoid space of chronic constrictive injury (CCI) rats. The expression of Fos protein in the dorsal horn of spinal cord was determined by immunohistochemistry. Results An immortalized rat astrocyte strain secreting enkephalin under the control of doxycycline was established successfully. After transplantation of IAST/TetOn/hPPE cell into the subarachnoid space of chronic constrictive injury (CCI) rats, the sensitivity of mechanical allodynia and the expression of Fos protein were significantly decreased (P<005), so the transplantation of IAST/Tet On/hPPE cell alleviated significantly CCIinduced chronic neuropathic pain in rats and the analgesic effect was also able to be regulated by Dox.Conclusion An immortalized rat astrocyte strain expressing enkephalin regulated by Dox has been established, which may provide a new tool for regulatable

Objective To explore the regulating mechanism of the astrocytes on the expression of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs) subunit in epileptogenesis. Methods The astrocytic conditional medium (ACM) was collected after being stimulated by glutamate, and then ACM was added to the cultured hippocampal neurons. The expression changes of neuronal GluR2 and protein interacting with C-kinase-1 (PICK1) mRNA were detected by RT-PCR. Results In the cultured hippocampal neurons, the GluR2 mRNA expression was significantly decreased at the 2nd, 8th, and 12th hours after the administration of ACM compared with that in the control group (P<0.05). However, the PICK1 mRNA expression was significantly increased at the same time points after the administration of ACM compared with that in the control group (P<0.05). The ionotropic glutamate receptor antagonists D-AP5 and CNQX could not fully block the action of ACM. Conclusion In epileptogenesis, the activation of astrocytes can downregulate the expression of neuronal AMPARs GluR2 subunit by upregul

Objective To study the expression of growth-arrest-specific protein 7 (Gas7) in the hippocampus of rats after chronic multiple-stress. Methods thirty-six Wistar rats were randomly divided into two groups: the chronic multiple stressed group and the control group. Rats in the stressed group were administered with four kinds of stressors including vertical rotation，sleeping deprivation，restraint(6hours/day) and illumination irregularly and alternately for 6 weeks. Then the expression of Gas7 protein in the hippocampus of rats was assayed by immunocytochemical method and Western blotting technique respectively and apoptotic cells were observed. Results The expression of Gas7 was extensively found in the hippocampus of both the control group and the experimental group mainly in the endochylema and processes of the neurons in the CA1 area. Positive staining, average absorbance in the CA1 and dentate gyrus of the stressed rats were higher than that in the control group (P<0.05). There was no difference in the expressions in the CA3 areas in the two groups (P>0.05). Caspase-3 positive cells were also obserued in the stressed rats. Western blotting detection indicated that the expression of Gas7 in the stressed rats was significantly higher than that of the control group(P<0.05).Conclusion Gas7 may participate in protecting neurons in the stess and promote the neurogenesis an development of neurons.

Objective To study the human lymphangiogenesis in the early stage of embryo and the expression of forkhead box C2(FOXC2) in human lymphangiogenesis. Methods Lymphatic vessel endothelial haluronic acid receptor-1(LYVE-1) was used as the special marker of lymphatic vessels.Immunohistochemical and double immunofluorescence staining were used to detect the expressions of FOXC2 and LYVE-1 in lymphatic vessels of 85 human embryos of 5 to 11 weeks pregnancy and analyze the features of lymphatic vessel genesis and development. Results LYVE-1 was initially detected in lymphatic vessels in 7-week human embryos. The lymphatic vessel endothelial cells presented brown staining for LYVE-1 in jugular and thoracic region. LYVE-1 was also found to express in human embryonic mesentery lymphatic vessels on the 70th day of pregnancy. The expression of FOXC2 in human embryo was detected prior to that of LYVE-1 At the beginning of the 6th week of pregnancy, FOXC2 was obviously seen in human embryonic mesoderm mesenchyme.FOXC2 expressed not only in human embryonic lymphatic vessels,but also in the vertebral body,cardiovascular wall and other tissues.The expressions of FOXC2 and LYVE-1 were still seen in human embryonic lymphatic vessel endothelial cells after 80 days of pregnancy. Conclusion Lymphangiogenesis of human embryo begins between the 7th and 8th weeks of pregnancy. FOXC2 and LYVE-1 could have some relation with the human embryonic lymphatic vessel genesis and development.

Objective To investigate the different expression patterns of the phenotypic marker protein for stromal cell, proliferating cell nuclear antigen(PCNA) and estrogen receptor α(ERα) in normal human prostate (NP) and benign prostatic hyperplasia (BPH) tissue, and to locate cells stained positively with the marker protein for stromal cell and PCNA or ERα at the same position. Methods Serial sections of 4 NP and 8 BPH specimens were subjected to immunohistochemistry (IHC) staining for vimentin, α-smooth muscle actin (α-SMA), myosin, PCNA and ERα. Results Compared with that in the NP tissues, the number of α-SMA positive cells increased significantly in BPH tissue, and the number of vimentin positive cells increased moderately in the stroma and prominently surrounding the acinus and in the basal layer. In BPH tissue, the myosin and ERα staining signal was lost in the stromal cells surrounding the acinus, and the positive staining cells gathered into bunches in the stroma far away from the acinus, while the positive cells were sporadically distributed in the NP tissue. The PCNA positive cells increased moderately in the stroma and increased significantly in the basal layer. The serial sections staining results showed that there was a colocation of PCNA, vimentin and ERα in basal cells, and a colocation of myosin and ERα in stromal cells. Conclusion There are apparent phenotypic modulations of the stromal cells in the BPH specimen co

Objective To investigate agerelated changes in rat white matter with the new stereological techniques in order to figure out the exact reason for the decline of myelinated nerve fibers in aged white matter and explore the effect of sex on the agerelated change of brain white matter. Methods Nine adult Long-Evans rats (6-8 months) and 8 aged Long-Evans rats (18 months) were used. The volume density, length density, total volume and total length of unmyelinated fibers in white matter were estimated with the stereological techniques and transmission electron microscope technique. Results In the male and female rats, there was no significant decrease in the total length of unmyelinated fibers in white matter of young and aged rats. There was a significant decrease in the total volume of unmyelinated fibers in white matter of aged female rats compared with that of young female rats. The total volume of unmyelinated fibers in white matter of aged male rats was decreased by 31% compared with that of young male rats even though the difference was not statistically significant. In young and old rats, there was no significant differences of all unmyelinated fiber parameters between male and female rats. Conclusion In aged rat white matter, there was a significant decrease in the unmyelinated fibers with large diameters. Meanwhile, the myelinolysis of the myelinated fibers with small diameters in aged white matter made the age-related decrease of the unmyelinated fibers with small diameters in white matter unnoticeable.

Objective To study the effect of the extract of total flavonoids of Chrysanthemum indicum (TFC) on adjuvant arthritis(AA) synoviocytes. Methods Totally 0.1ml of the complete Freund’s adjuvant was subcutaneously injected into the right hind feet pads of 20 SD rats. 24 days after immunity synoviocytes in the knee joint were treated with TFC. Cell morphology was examined with electron microscopy. Protein level of caspase-3 cleaved fragments was analyzed by Western blotting. The annexin V stain assay was applied to explore the effect of caspase-3 inhibitor on the amelioration in synovial cells apoptosis of AA rats. Results Typical morphology and biochemical feature of apoptosis in synovial cells of AA rats were observed with TFC. The protein level of caspase-3 cleaved fragments increased obviously and was related with the concentration of TFC in synovial cells of AA rats. The apoptotic cells positively stained with annexin V were markedly reduced by caspase-3 inhibitor. Conclusion TFC can induce apoptosis in AA rats synoviocytes，which may achieve therapeutical effects in AA. The activation of caspase-3 may

Objective To establish an ideal focal cerebral ischemia reperfusion model in mI>onkey/I>s. Methods Adult healthy rhesus mI>onkey/I>s (Macaca mulatta) 12 cases (male 6 and female 6), aged 4-7 years and weighted 4.8-7.5kg. were used in this study. The middle cerebral artery occlusion/reperfusion (MCAO/R) model was established by inserting a standard micro-balloon catheter intraluminally from the carotid common artery or femoral artery into the proximal segment of the middle cerebral artery (MCA). The regional cerebral blood flow of MCA was occluded by expanding the micro-balloon to cause ischemia, and withdrawing the micro-ballon catheter to reperfuse the MCA. The MCAO/R model was evaluated by angiography, magnetic resonance angiography (MRA), magnetic resonance imaging(MRI), tetrazolium chloride (TTC) staining and neurological behavoral function scores. Results By inserting a micro-balloon catheter intraluminally from the carotid common artery or femoral artery into the MCA, the micro-balloon catheter could be inserted into the MCA to occlude blood flow, and no image of MCA shown on TV screen. In MCA blood flow supplied area, magnetic resonance T1, T2 and DWI showed high signals, TTC staining showed cerebral ischemic infarction, and correspondly the mI>onkey/I>s showed neurological function disorders. This method used a simple operatire procedure had a high successful rate, and could be repeated. Conclusion We showed ideal method to establish the MCAO/R model in mI>onkey/I>s by inserting intraluminally a micro-balloon catheter into the MCA.

Objective To study the repairing effects of nerve growth granule (NGG) on rat common peroneal nerve transection injury. Methods After 50 Sprague-Dawley rats were subjected to nerve suture after transaction, they were randomly divided into 5 groups for daily intragastric administration of drugs: NGG high-dose (5.2g/kg), medium-dose (2.6g/kg), low-dose (1.3g/kg) groups, mecobalamin group (positive control) at 625 μg/kg, control group (control group control). The drug administration lasted for 4 weeks. Footprint test was performed 2-, 3- and 4- weeks after surgery to evaluate toe spread function (TSF). Electrophysiology was performed 4 weeks after operation to determine the compound muscle action potential (CMAP) and nerve action potential (NAP). The number of regenerated myelinated nerve fibers, thickness of myelin sheath and cross sectional area of tibial muscle were measured by histomorphology. Results TSF, amplitude and recovery rate of CMAP and NAP, the number of regenerated myelinated nerve fibers, thickness of myelin sheath and section area of tibial muscle were all increased significantly in a dose-dependent manner compared with the control group. Conclusion NGG contributes to axon growth and myelination, and thus promotes peripheral nerve regeneration in rats with functional recovery.

Objective To observe the morphologic changes of immature dendritic cells(imDCs) before and after the electransfection of human hepatic cancer cell RNA. Methods Monocytes were purified from human peripheral blood, and induced into imDCs. Then human hepatic cancer cell RNA was electransfected into monocytederived imDCs. ImDCs were identified by the immunocytochemical method with 7 specific antibodies before and after electransfection. These dendritic cells were observed by scanning electron microscopy. Results After electransfection of human hepatic cancer cell RNA there were few changes of molecule expressions in imDCs. ImDCs were in round, oval and irregular shapes before electransfection. Their sizes were not identical but all bigger than monocytes. There were many protrusions in different shapes which looked like dendrite, or/and bubble, veil cloud on the surface of these imDCs. Although there were some cell fusions and cell deaths after electransfection, most imDCs recovered from the damage. Electransfecting human hepatic cancer cell RNA into imDCs would make pores on cell membrane.Conclusions The pores on cell membrane make it possible that the exogenous material enters imDCs. This study can prove the possibility of electransfecting human hepatic cancer cell RNA into imDCs to make cancer vaccine, which provides a new way for tumor biologic therapy.

P>Objective Insulin resistance is a possible cause of polycystic ovary syndrome (PCOS). It is presumed that ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) is associated with insulin resistance. The purpose of this study is to explore the expression of ENPP1 in granulosa cells of the ovary and its relationship with PCOS. Methods Twelve aliquots of follicular granulosa cells were isolated from 12 samples of the patients with PCOS and 22 aliquots from 22 samples of the patients without PCOS, respectively．ENPP1 expression was analyzed by reverse transcription polymerase chain reaction (RT-PCR ) and in situ hybridization. The relative expression of ENPP1 mRNA was detected by realtime quantitative polymerase chain reaction (realtime PCR). Results ENPP1 was expressed in granulosa cells of the ovary. ENPP1 expres

Objective To study the significance of the expression of mitochondrial DNA Cyt-b and vascular endothelial growth factor mRNA in human breast cancer. Methods Reversedtranscription polymerase chain reaction（RT-PCR）was applied to detect the expressions of mtDNA Cyt-b and VEGF in 19 samples of breast cancer tissue and 19 samples of peritumoral tissues. β-actin served as a quantitative standard marker. The different expressions in breast cancer tissues and paracancerous tissues were compared and their relations with clinical pathology parameters were analyzed. Results The expressions of mtDNA Cyt-b and VEGF mRNA in 19 samples of breast cancer tissues were higher than those in paracancerous tissues, with a significant difference between the two groups (P<0.01). The expression of mtDNA Cyt-b was positively correlated with the expression of VEGF mRNA (P<0.01) and the expression of VEGF145 and VEGF189 mRNA (all P<0.05). The expression of VEGF145 and VEGF189 mRNA, which were the isomerides of VEGF, was higher in breast cancer tissues than in paracancerous tissues, with a difference between the two groups (P<0.05). The expression of mtDNA Cyt-b and VEGF mRNA in lymph node metastasis group was higher than that of nonlymph node metastasis group, with a difference between the two groups (P<0.05). The expressions of mtDNA Cyt-b and VEGF mRNA in phase Ⅰof TNM stage were lower than those in phase Ⅱ, with a difference between the two groups (P<0.05). There was a significant differe

Objective In order to make further investigation of the functional meaning of Ca2+dependent phospholipids binding protein AnnexinⅡ, we tested the effects of AnnexinⅡ combined with embryonic neural stem cells (NSCs) transplantion on the repair of spinal cord injury (SCI). Methods The spinal cord of the adult rats was transected completely between T9T10. AnnexinⅡ and NSCs were injected at the transected site. The lesion area of the spinal cord, growth of axon, and survival number and migration of the transplanted NSCs were measured. The survival number was shown by prelabeling the NSCs with Hoechest 33342 and the growth of axon traversing the transected area was shown by fluorogold (FG) retrograde tracing. The AnnexinⅡ injection+NSCs implantation groups were compared with groups that respectively received 1. NSCs implantation alone; 2. shamoperation+NSCs implantation and 3. Vehicle injection of culture medium. Results Our results demonstrated that AnnexinⅡ treatment in vivo could significantly reduce the lesion area (P<001), increase the number of FG retrogradelylabeled neurons rostral to the injury (P<001), and improve the

Objective To study the effect and significance of hyperbaric oxygen treatment on the expression of neuroglobin by observing the changing tendency of neuroglobin in mouse hippocampus after forebrain ischemiareperfusion. Methods Eighteen female C57BL/6N mice were divided into three groups randomly: the shamoperated control group, the 48hour ischemiareperfusion group (48h I/R) and the group treated with hyperbaric oxygen after 48hour ischemiareperfusion (48h HBO). HE staining and immunohistochemistry experiments were carried out on the brains of all the animals. Image analysis and statistical treatment were also performed for all the images. Results The neuroglobin expression level of 48h I/R group and the 48h HBO group were higher than that of the shamoperated group (P<0.01 and P<0.001, respectively); the optical density of the 48h HBO group was higher than that of the 48h I/R group(P<0.01).Conclusion Hyperbaric oxygen treatment had an obvious effect on ischemic brain injury, and increasing t

Objective Recent researches showed that Eph/Ephrin tyrosine kinase family plays an important role in the development and functional maintanence of nervous system, but it is unclear if it affects sympathetic nervous system.The study is to detect the effect of EphB/Ephrin-B1 signal on the currents of nicotinic acetylcholine receptor (α7-nAChRs) on ciliary ganglion neurons. Methods Currents of α7-nAChRs on cultured and acute-associated ciliary ganglion neurons were recorded respectively. Cells were randomly divided into four groups: the control group, the EphrinB1Fc treated group (cells stimulated with different concentrations of Ephrin-B1Fc which is a recombination of IgG and Ephrin-B1), IgG treated group (cells stimulated with IgG at the same concentration of IgG needed for Ephrin-B1Fc recombination), Ephrin-B1 treated group (cells stimulated with Ephrin-B1 at the same concentration of Ephrin-B1 needed for Ephrin-B1Fc recombination). Results There was no difference between the control, IgG and Ephrin-B1 groups, but Ephrin-B1Fc could inhibit α7-nAChRs currents of both cultured and acute-associated CG neurons in a concentration-dependent manner. The inhibition happened quickly and was reversible. ICSUB>50/SUB> of ISUB>peak/SUB>/CSUB>m/SUB> of cultured and acuteassociated cells was 0.032mg/L and 1.4mg/L respectively. Conclusion The results showed that EphB/Ephrin-B1 signal can affect currents of CG α7-nAChRs. Is suggests that

Objective To explore the relationship between the expression of BMP-7 in the hepatic tissue and hepatic fibrosis in rat hepatic fibrosis induced by CCl-4. Methods Healthy adult male Wistar rats were randomly divided into the control group and the hepatic fibrosis group. The control group was subcutaneously injected with oily solution of 0.12ml per 100 grams of rat weight every monday and thursday; the hepatic fibrosis group was subcutaneously injected with 60% carbon tetrachloride oily solution of 0.3ml per 100 grams of rat weight every monday and thursday. Rats were continuously injected for 6 weeks, 10 weeks, 16 weeks and 21weeks respectively. When the experiment ended, all the rats were killed. The hepatic middle lobule was taken and embedded with paraffin. Collagen fibers in the hepatic tissue were detected by Sirius red staining. The expression of BMP-7 in the hepatic tissue was detected by immunohistochemical staining and Western blotting. Results In the rat hepatic tissue, there were few collagen fibers in portal veins and the portal area in the control group. Proliferation of collagen fibers could be observed in each rat of the hepatic fibrosis group and hepatic fibrosis was aggravated with the lasting of the experiment. In immunohistochemical staining, several BMP-7 positive cells were observed in the rat livers of the control group; many BMP-7 positive cells were observed in the 6-week group. The expression of BMP-7 positive cells gradually decreased after 10 weeks,and they were seldom seen in the 21-week group. The Western blotting results were basically the same as those of immunohistochemical staining.Conclusion In rat hepatic fibrosis induced by carbon tetrachloride, the expression of BMP-7 in the hepatic tissue was opposite to that in the hepatic fibrosis. BMP-7 can protect the hepatic fibrosis to some degree.

Objective To determine the effects of nerve regeneration factor (NRF) on neurite growth in cultured hippocampal neurons of rats. Methods After different periods (6h, 12h, 24h) of incubation by different concentrations (0.25mg/L, 0.5mg/L, 1.0mg/L) of NRF, the hippocampal neurons were observed and photographed by phase contrast microscopy, and the neurite length was analyzed using the Scion software. Realtime fluorescence quantitative RT-PCR was performed to examine the mRNA levels of growth associated protein 43 (GAP-43) after different incubation periods with different concentrations of NRF. Fluorescent immunocytochemistry and Western blotting were performed to further examine the protein levels of GAP-43 in cultured hippocampal neurons after 24h incubation with different concentrations of NRF. Results NRF promoted the neurite growth of cultured hippocampal neurons with the most obvious mophological changes appearing upon treatment with NRF at 1mg/L for 24 hours. Realtime fluorescence quantitative RT-PCR showed that the mRNA level of GAP-43 in cultured hippocampal neurons was upregulated by NRF in a dose and timedependent manner, with the best concentration of 1mg/L. Meanwhile, fluorescent immunocytochemistry and Western blotting showed that the protein level of GAP-43 was upregulated by NRF with the best concentration of 1mg/L. Conclusion NRF could promote the neurite growth and modulate the expression of GAP-43 in cultured hippocampal

Objective To explore the evidence for the morphologic comparability between the meridians and interstitial connective tissues in the lower limb with automarked and 3DRebuilding technology. Methods First, the muscles were extracted and noise was eliminated; With edge detection a convex hull of the muscles edge was found; The interstitial connective tissue points were marked in the cross section images of left leg from the digital dataset of Virtual Chinese HumanMale1 (VCH-M1). Finally, the automarked images were reconstructed with 3D-Doctor. Results The reconstructed meridians in the lower limb automarked with computer procedures tallied basically with the meridians and collaterals recorded in Traditional Chinese Medicine(TCM) illustrative graphics. Conclusion Having the advantages of digital anatomic study on acupuncture points earlier and mature 3D-Rebuilding techniques,the department of Anatomy of South Medical University cooperated with the Department of Scientific Calculation and Computer Application of Sun Yat_sen University to devise a set of computer programs which could be used to AutoMark interstitial connective tissues in Virtual Human lower limbs and then reconstruct virtual fascia meridians which corresponded with the TCM records. This method avoided manmade error to a certain degree in the process of recognizing and rebuilding of fascias. The study is significant to explore the morphologic comparability between the meridians and col

钙离子（Ca²⁺ ）是通过局部信号获得特异性刺激的关键细胞内信使分子。Ca²⁺ 结合蛋白，如钙调蛋白（CaM）及其靶蛋白是Ca²⁺ 依赖性反应信号传导的关键靶点。钙/钙调蛋白依赖性蛋白酶Ⅱ（CaMKⅡ）是一种多聚体酶，它是哺乳动物前脑总蛋白的重要组成部分，并且是形成突触后致密部的主要成分。近年来国内外研究显示，CaMKⅡ包含α、β、γ 和 δ 4种亚型，其中α 和 β 主要在神经组织中表达，而 γ 和 δ 则在全身多种组织均有表达，它们参与特定的突触可塑性和记忆巩固过程，对神经系统的兴奋性及一些神经系统疾病的发生起重要作用。前期也有研究表明CaMKⅡδ在促进神经元存活中起重要作用。本文就CaMKⅡ的结构及其在神经系统中的作用和与相关神经系统疾病的关系作一综述。

Objective To analyze pulmonary blood supply in congenital heart disease with tetralogy of Fallot (TOF) and pulmonary atresia in young children. Methods The clinic and imaging data of 94 cases of congenital heart disease with tetralogy of Fallot (TOF) and pulmonary atresia were analyzed, aged 2-26 (11.25±8.24) months, from January 2004 to January 2007 Results There were 51 cases with pulmonary blood flow only from ventricle, 27 cases with pulmonary blood flow only from patent ductus arteriosus (PDA) and/or aortopulmary collateral arteries (APCAs), and 16 cases with pulmonary blood flow from both pulmonary and systemic arterier. There were no significant differences in the incidence of PDA and the degree of pulmonary stenosis (P>005). The differences were significant between the incidence of APCAs and the degree of pulmonary stenosis (P<0.05). Conclusion The source of pulmonary blood in congenital heart disease with TOF and pulmonary atresia is complex. The incidence of aortopulmonar