Table of Content

20 July 2009, Volume 29 Issue 7

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研究论文

Reconstructions of neuron synapses involved in the function reestablishments of neuroimmunoreglation network of rats

Yang GAO; Xiao-tian XU; Xiang-qian KONG; Li-chuan HONG; Huo-yuan LIU; Jia-you LIN

2009, 29(7):  673-677. 

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Objective: This study was to explore the function reestablishment patterns of neuroimmunoregulation network. Methods: Whole-some gene array was performed to analyze the genes expression difference involved in neuroimmunoregulation function of the lateral hypothalamic (LH) in rats at different immunizational time. The function trees of different genes were analysed with the PathWay Miner public database. Results: 632 of the genes were considered to be differentially expressed in comparison to the control group: 374 of 2-day immunized group，62 of 4-day immunized group ，and 196 of 6-day immunized group. Function signal pathway analysis for 398 up-regulated genes shows 27 genes evolved in 31 cell functional signal conduction pathways, including the well-known signal conduction pathways of synaptic reconstruction. Conclusions: Synapses reconstruction appears to be the important pattern of function reestablishments in neuroimmunoregulation network.

PP60C-SRC/ERK1/2 regulates the viability of rat spermatogonial stem cells in vitro

Jia-xiang CHEN; Si-fan XU; Jing-lei WANG; Jun-ling YANG; Hai-yan QIN; Ting ZOU

2009, 29(7):  678-682. 

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Objective The present study aimed to investigate the effect of PP60C-SRC on the viability of rat spermatogonial stem cells in 9 day-old rat. Methods MTT method was used to observe the viability of the spermatogonial stem cells treated with antisense C-SRC oligodeoxynucleotides (ODNs) and PD98059 in vitro; Western blot was used to observe the PP60C-SRC and phosphorylated extracellular regulated kinase1/2 (p-ERK1/2) contents. Results Compared with that in control group, the viability of the cells decreased by 8.1 % (P<0.05) and 9.4 %(P＜0.01), respectively, after the spermatogonial stem cells were treated with antisense C-SRC ODNs and PD98059. Compared with the control group, the PP60C-SRC and p-ERK1/2 contents decreased by 33.8 % and 38.4 % (both P<0.01), respectively, in the spermatogonial stem cells after transfected antisense C-SRC ODNs. Compared with the control group, the p-ERK1/2 contents decreased by 34.5 % (P<0.01) in the spermatogonial stem cells after treated with PD98059. Conclusion PP60C-SRC regulates the viability of rat spermatogonia stem cells through p-ERK1/2.

Prossible mechanism of Nidus Vespae protein (1) inhibits proliferation of bronchial smooth muscle cell of rats

Chang-dong WANG; Peng CHEN; Xu YAN; Guang-yuan HE

2009, 29(7):  683-687. 

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Objective NVP (1) was one pure protein that isolated from Nidus Vespae. We found that NVP (1) can inhibit proliferation of bronchial smooth muscle cell. At the same time, we investigated the mechanism of NVP (1) in inhibiting proliferation of bronchial smooth muscle cell. Method Primary cell culture. The effects of NVP (1) on bronchial smooth muscle cells proliferation by MTT assay. The cell cycle was detected by flow cytometry. Western-blotting was used to reveal p21, cdk2 protein expression. Results NVP(1) (6.4×10-3g/L) can arrest bronchial smooth muscle cell in cycle G1 and increase the expression level of p21. On the contrary, NVP (1) can inhibit proliferation of bronchial smooth muscle cell by down-regulating cdk2 protein expression. Conclusion NVP (1) inhibits proliferation of bronchial smooth muscle cells and induces G1 cell cycle arrest.

Analysis of Proteomics of Seminal Plasma from Idiopathic Male Infertility

Xu-yi REN; Min-lan ZHOU; Xin-zong ZHANG; Yang LIU; Run-liu YU

2009, 29(7):  688-690. 

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Objective To discover biological markers. Methods Two-dimensional electrophoresis and Mass-Spectrum techniques (MS+MS/MS) were used for analyzing the seminal plasma from idiopathic male infertility and control. Results Serum albumin and prostatic acid phosphatase were reduced in seminal plasma from idiopathic male infertility rather than the control, while Cathepsin B and Zn-alpha-2-glycoprotein were increased. Conclusion Idiopathic male infertility possibly was associated with disorder of sperm capacitation and seminal immune function.

Increased phosphorylation of p38 MAPK participated in the attenuation of MCAO-induced brain injuries by hypoxic preconditioning of mice

Shu-jun JIANG; Nan ZHANG; Xiang-ning BU; Xu LIU; Su-juan FENG; Jun JIANG; Wei-wei YANG; Jun-fa LI

2009, 29(7):  691-696. 

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Objective　To explore the effect of hypoxic preconditioning (HPC) on middle cerebral artery occlusion (MCAO)-induced brain injury of mice and the changes of p38 mitogen activated protein kinase (p38 MAPK) phosphorylation and protein expression levels in the ischemic cortex. Methods　Using our established HPC and MCAO mouse models, healthy male BALB/c mice weighing at 18～20 g were randomly divided into 4 groups: H0 sham, H4 sham, H0 and H4 group. The brain infarct volume and neural injury was determined by 2,3,5-Triphenyltetrazolium chloride (TTC) and Nissl staining. Western blot combined with GelDoc imagine systems were applied to examine the changes in p38 MAPK phosphorylation and protein expression levels in the brain of mice. Results　HPC significantly attenuate the brain injury induced by MCAO (P<0.05). Phosphorylation levels of p38 MAPK in the ischemic core and penumbra increased significantly when compared with H0 sham group (P < 0.05, n = 6 for each group). HPC increased the phosphorylation levels of p38 MAPK in the penumbra and contralateral cortex (P<0.05, n=6). However, there were no significant changes in p38 MAPK protein expression levels. Conclusion p38 MAPK might involved in the attenuation of MCAO-induced brain injury by HPC of mice.

Therapeutic effects of HLA A silencing and TH over-expressing human fibroblasts grafted in a rat model of Parkinson's disease

Cai-xia LIANG; Yun-zhi XU; Chun-li ZHAO; De-yu ZHENG; De-yi DUAN

2009, 29(7):  697-703. 

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Objective Human leukocyte antigen A (HLA A) silencing and tyrosine hydroxylase (TH) over-expressing human embryonic lung fibroblasts (HELFs) were grafted into the striatum of a rat model of Parkinson's disease (PD) to observe rotation behavior of the PD rats and inflammation in the brain. Methods The cultured HELFs were transduced with retrovirus carrying TH and lentivirus harboring HLA A siRNA and green fluorescent protein (GFP), and then grafted into the striatum of PD rats. The rotation behavior was measured, immunostaining for TH, OX42, OX6 and GFAP performed, and GFP was used to evaluate the survival of the grafted cells. HELFs and, HELFs and rat embryonic lung fibroblasts transduced with TH-containing retrovirus were grafted into the brain to serve as controls. Results The GFP fluorenscent cells were gradually reduced after transplantation, and relatively more cells were found in the HLA A silencing HELFs without significant amelioration of rotation behavior. The number of OX42-positive microglia was observed unchangeable, while OX6 cells were gradually increased. Activation of GFAP-positive astrocytes decreased with time, but significantly stronger than that in the contralateral side. Conclusions HLA A silencing could not promote amelioration of rotation symptom and intracerebral transplantation could induce activation of microglia and astrocytes.

TSA Promotes Mesenchymal Stem Cells differentiating into cardiomyocyte-like cells

Li-li ZHAO; Jing ZHU; Chuan FENG; Ge YANG; Jie TIAN

2009, 29(7):  704-708. 

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Objective To observe the effect of TSA to induce mesenchymal stem cells(MSCs) differentiating into cardiomyocyte cells, to explore the effective conditions． Methods MSCs were incubated in different concentration of TSA and different duration time, then detected the histone deacetylase(HDAC) activity; the differentiation of MSCs was observed by phase-contrast mictmcopy;fluorescence immunohistochemistry assessed cardiac troponin T and RT-PCR detected the expression of Nkx-2.5、GATA-4 genes．Result The HDAC activity was the lowest when incubated with 300nmol/L TSA for 24 hours, expression of cTnT of the cells was enhanced after incubated with TSA. The expression levels of Nkx-2.5、GATA-4 genes were enhanced before interference.Conclusion TSA can advance MSCs to differentiate into cardiomyocyte cells in vitro in a suitable condition.

Culture of cynomolgus monkey embryonic stem cells and differentiation to neural lineages

Wan-wan ZHU; Di WU; Ning LI; Chun-lin ZOU; Yan-ling XU; Yun-qian GUAN; Yu ZHANG

2009, 29(7):  709-715. 

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Objective Establish a culture method and a differentiation method for the cynomolgus monkey ES cells, to prepare for future experiments of transplantating neural progenitors derived from cynomolgus monkey ES cells into monkey disease model. Methods Prepare mouse embryonic fibroblasts (MEF) as feeder cells, culture the cynomolgus monkey ES cells for a long period, then characterize the cells by marker staining. The cynomolgus ES cells were differentiated into neural progenitors by adding noggin into differentiation medium, and the cells were stained in each stage during differentiation by immno-histochemistry. Results The cynomolgus monkey ES cells form colonies, and express cell markers of undifferentiated primate ES cells for more than 20 passage doublings. In neural differentiaion assay, the ES cells formed rosette and neural tube after 14 days neural induction. The staining showed large numbers of Nestin positive and Tuj-1 positive neural progenitors. If induction period extended to 21 days, the number of Nestin positive and Tuj-1 positive cells was still numerous. After 35 days of differentiation, many GFAP positive cells started to emerge, and Tuj-1 positive cells decreased. Conclusions The culture system of cynomolgus monkey ES cells was successfully established. The induction of ES cells toward neural lineages produced a large number of neural progenitors, and non-mature neurons expressing Tuj-1 marker.

The role of TRAF6 in NF- B and AP-1 signaling transduction pathway in human B lymphoma cell line

Wen ZHANG; Xuan ZHANG; Xiao-feng ZENG; Feng-chun ZHANG

2009, 29(7):  716-720. 

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Purpose To investigate the role of TRAF6 in NF- B and AP-1 signaling pathway in human B cell line. Methods Human Ramos B cells were transfected with plasmids expressing YFP fusion dominant-negative TRAF6 (DN-TRAF6), or transfected with shRNA-TRAF6 plasmid. After cultured overnight, cells were either sorted with flowcytometry or screened by G418. Activation of NF- B and AP-1 pathway, including phosphorylation of I B , ERK, JNK and P38, as well as nuclear translocation of NF- B subunits (P65, P50 and c-Rel)and AP-1 subunits(c-Fos, c-Jun, CREB and ATF) were detected by Western blot and ELISA. Results In cells which overexpress DN-TRAF6 or endogenous TRAF6 expression were knocked-down by shRNA, the phosphorylation of I B , as well as phosphorylation of JNK were inhibited. Furthermore, nuclear translocation of NF- B subunits P65, P50, c-Rel,and AP-1 subunits c-fos and c-jun were also inhibited in B cells overexpression of DN-TRAF6. Conclusion TRAF6 selectively activates some kinases in CD40 mediated NF- B and AP-1 signaling pathway, and plays an important role in their activation.

New Model -Track of β-cell Development By Generation of Transgenic Zebrafish

Ming XIA; Xue PAN; Min DENG; Yi JIN; Yi CHEN; He-sheng WANG; De-ming KONG

2009, 29(7):  721-725. 

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Objective we have established pancreatic β-cell developmet fish model, which showed specific spatial expression patterns in vivo. Methods molecular clone, microinjection, whole embryo in-situ hybridization（WISH）and fluorescence microscopy in living were used to analyze of β-cell development by generation of transgenic zebrafish. Results screening and established pancreatic β cells of transgenic zebrafish, then confirmed that fluorescence protein expression in the same spatiotemporal pattern with endogenous insulin gene to achieve dynamic monitoring islet β-cell development situation in vivo. Conclusion the pancreatic β cells of transgenic zebrafish animal model can be successfully traced pancreatic β cell development in whole level.

Over-expression of NOTCH-1 inhibits TNFα-induced apoptosis and proliferation suppression in human gastric carcinoma cells

Jun YAO; Cui-juan QIAN

2009, 29(7):  726-730. 

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Objective To explore the regulative function of over-expression of NOTCH-1 in TNFα-induced apoptosis of gastric carcinoma carcinoma BGC-823 cells. Methods BGC-823 cells were infected with retrovirus recombined with intracellular domain of NOTCH-1.The cell clones, into which objective genes had been transfected, were obtained after being screened with G-418. Before and after incubated with TNFα, MTT assay, flow cytometry,Western blot and EMSA were used to detect cell growth, apoptosis, expressions of NOTCH-1, NF-κB and caspase-3 activity. Results There was stronger expression of NOTCH-1 and its target gene HES-1 in the cells transfected with NOTCH-1 gene than that of control. After TNFα treatment, the rates of killing rates and apoptosis in NOTCH-1-transfected cells were all lower than control cells. Moreover, we showed over-expression of NOTCH-1 suppressed caspase-3 activation induced by TNFα. However, the TNFα-induced activation of NF-κB was not affected by over-expression of NOTCH-1. Conclusions over-expression of NOTCH-1 significantly protects BGC-823 cells from apoptosis and growth suppression induced by TNFα. This effect is mediated, at least in part, through inhibition of caspase-3 activation independent of NF-κB.

Human bone morphogenetic protein 3(hBMP3) inhibited the growth of human osteosarcoma cells MG63 and U2OS in vitro

Li-mei WU; Huan-ling HE; Xing-xing LI; Ying-hua CHEN; Jia WEI; Guo-wei ZUO; Jing-kun MIAO; Yan WANG; Lan ZHOU

2009, 29(7):  731-736. 

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Objective The biological effects of BMP3 on osteosarcoma were investigated by treating human osteosarcoma cell lines MG63, and U2OS with human BMP3 (hBMP3). Methods Osteosarcoma cells in experimental groups were respectively treated with AdBMP-3 and rhBMP3-CM, control groups with AdGFP and rGFP-CM, the blank with neither. The abilities of their proliferation, apoptosis, transmigration and differentiation were respectively detected by trypan blue exclusion test, terminal deoynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) and acridine orange-ethidium bromide fluorescent stain (AO/EB), transwell-room and alkaline phosphatase(ALP) activity reagent kit. Results 1) All the indexes detected were not significantly different between two control groups. 2) Compared with control groups, the cell survival rates showed a significant decrease in experimental groups. 3) The apoptosis indexes increased. 4)The number of trans-membrane cell decreased 5) The activity of alkaline phosphatase increased after treating with AdBMP3 and rhBMP-3 for 3 days in MG63, 5 days in U2OS. Conclusions hBMP3 could inhibit osteosarcoma cells and promote bone formation.

Establishment and characterization of a mouse medulloblastoma cell line

Yue-yuan WANG; Yun-zhou GAO; Zhao-qi WANG; Yan SHEN; Wei-min TONG

2009, 29(7):  737-741. 

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Objective In order to study the molecular mechanism of medulloblastoma, the cell line with PARP-1 and p53 null mutation was established and characterized. Method Mouse medulloblastoma cell line derived from PARP-1/p53 double knockout mice was established. We analyzed cell characters after 30 passages, using neuronal cell-specific markers by immunofluorescence. The cells were transfected with pEGFP-C1-Hparp-1 and pEGFP-C1 plasmids，and the expression of PARP-1 protein was detected by immunofluorescence stainning and Western blot. Result The cells showed positive immunoactivity for the neuronal-specific markers such as Vimentin, Dcx and III-Tubulin, and cells were negative for PARP-1 protein. Exogenous PARP-1 expression was visualized by immunofluorescence and Western blot analysis after pEGFP-C1-Hparp-1 transfection. Conclusions Mouse medulloblastoma cell line with defective DNA damage response has been successfully established, and provides a useful tool for further dissecting the molecular mechanism and pathogenesis of medulloblastoma.

Genetic polymorphisms of seventeen Y-chromosomeal STR loci in Heiyi Zhuang ethnic groups in Guangxi province

Shao-kang TENG; Lin-zhi CAO; Shi-ning HUANG; Chang-sheng HUANG

2009, 29(7):  742-748. 

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Objective To investigate the Allelic and haplotype frequency distribution of seventeen short tandem repeat loci of Y chromosome in Heiyi Zhuang ethnic groups in Guangxi province. Methods Seventeen Y-STR loci , of which the template DNAs were extracted from blood samples of 184 unrelated male individuals in Heiyi Zhuang population, were amplified by using the AmpFlSTR? Yfiler?. The PCR products were genotyped with ABI PRISM 310 genetic analyzer. Results 　 The Gene diversity ranged from 0. 4910 to 0.9727 at DYS456、DYS389Ⅰ、DYS390、DYS389Ⅱ、DYS458、DYS19、DYS385a\b、DYS393、DYS391、DYS439、DYS635、DYS392、Y-GATA-H4、DYS437、DYS438、DYS448 . A total of 180 different haplotypes were observed , The haplotype diversity value calculated from all 17 loci combined was 0. 99976. The significant difference of the allelic frequency distribution in Y-STR loci was observed between Heiyi Zhuang population and other observed populations. Conclusion 　The 17 Y-STR loci in Heiyi Zhuang population of Guangxi province are highly affluent genetic polymorphic and can offer valuable genetic datas for paternity testing and paternal genetic lineages evolution.

Effects of Androgen on Mouse Model of Autoimmune Ovary Failure

Xiao-bo SHI; Na LI; Qiu-hua LIN; Can LIAO; Li-qing FAN

2009, 29(7):  749-752. 

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Objective:To identify the effects of androgen on mouse model of autoimmune ovary failure. Methods: Fifty mice were chosen and randomly divided into five groups of ten, that were the control group, early therapeutic group and model group, late therapeutic group and model group. Autoimmune POF was induced in forty mouse by Pzp3. Then, animals were randomly divided into four groups of ten; two of the groups were treated with androgen from the 15th and 20th day after immune. The control group was treated with distilled water. The therapeutic efficacy of each treatment was evaluated by observing and measuring the changes in sexual cycles of mice, the serum level of AzpAb, infiltration of cells positively expressing CD45 in the ovary as well as the percentage of growing follicles of the ovary. Results: The sexual cycles, serum level of AzpAb, infiltration of cells positively expressing CD45 in the ovary of each model group was higher than control group; early therapeutic group was lower than model group; and serum level of AzpAb in early therapeutic group was lower than late therapeutic group. The percentage of growing follicles in the ovary of each model group was lower than control group; early therapeutic group was higher than model group and late therapeutic group. Conclusion: The use of androgen in mice with autoimmune POF could notably ameliorate the pathogenetic conditions of the disorder, and the treatment during prophase was apparently more effective than during the advanced stage.

临床园地

Diagnosis and Treatment of Subclinical Pheochromocytoma

An-li TONG; Zheng-pei ZENG; Ming LI; Shi CHEN; Lin LU; Di YANG; Han-zhong LI

2009, 29(7):  753-756. 

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Objective To analyze the clinical features of patients with subclinical pheochromocytoma (PHEO). Methods Review of clinical features of 22 patients with subclinical PHEO treated in PUMC hospital from 1997 to 2007. Results All patients were asymptomatic. 24hrs urinary catecholamine excretion was detected normal in 10 of 22 cases, while increased in the others. 16 patients were prepared withα-receptor blocker before operation. During the operation, BPmax (maximal blood pressure) before tumor resection, BPmin (minimal blood pressure) after resection and ΔBP(BPmax－BPmin) were 163±34/86±20 mmHg，105±12/61 ±10 mmHg and 58±37/25±21 mmHg, Respectively, in the prepared group. And they were 169±36/104±20 mmHg, 97±18/56±13 mmHg and 71±48/48±29 mmHg in the other 6 cases without α- blocker preparation. DBPmax and ΔDBP in the prepared group were significantly lower than the unprepared group. Conclusions Most patients with subclinical PHEO have increased catecholamine secretion. Blood pressure is fluctuant greatly during operation in some patients. Patients should be treated with α-receptor blocker preoperatively in order to decrease the operation risk.

研究短文

Phenoxybenzamine attenuates norepinephrine fluorescent intensity of iris in hemorrhagic shock rats

Geng ZHANG; Jun-xu REN; Zi-gang ZHAO; Chun-yu NIU

2009, 29(7):  757-758. 

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Changes of plasma angiotensin II level and expression of vessel PDGF-B in rats with carotid artery stenosis

Yin-ping SUN; Xing WANG; Da-wei ZHANG; Xiao-bing WANG; Yong GUO

2009, 29(7):  759-760. 

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Relationship Between Systemic Lupus Erythematosus and Epstein-Barr Virus Infection

Zhen-jun SHI; Tao WANG; Yue-hua LIU; Qin SI

2009, 29(7):  761-762. 

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Analysis of 23 cases of primary empty sella syndrome

Jun-yuan WANG; Zhao-hui LV; Ju-ming LU

2009, 29(7):  763-764. 

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Pentoxifylline ease of flap ischemia reperfusion injury in rats

Rong-kui GUO; Xin-ming WANG; Xi-an DONG

2009, 29(7):  765-766. 

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短篇综述

Regulatory mechanisms of PTEN

Jun-bo HONG; Nong-hua LV

2009, 29(7):  767-770. 

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PTEN (Phosphatase and Tensin Homology deleted from Chromosome ten)，a putative antioncogene,encodes a PTEN protein that has a dual specificity phosphatase activity with both protein phosphatase and lipid phosphatase，which is capable of regulating lots of molecules through inhibiting the PI3K/Akt signaling pathway.Recently，some scholars dedicate to the modulation of PTEN，which demonstrates that PTEN can be regulated by multiple factors in the level of gene and protein.This review briefly summarizes current knowledge about the modulation of PTEN and its mechanisms.

Influence of exercise on meal size and body weight

Jun WANG

2009, 29(7):  771-774. 

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There are several different points about the influence of exercise on appetite，meal size and body weight. Exercise affects body weight through two methods: energy expenditure and the influence on appetite and meal size. This article reviews that acute exercise has no effect on appetite and meal size; Long time exercise increases meal size of lean subjects and has no effect on overweight and obesity. These will be useful to weight loss on nationwide body-building.

医学教育

Construction of a patient-centered hospital information management system

Yuan ZHANG

2009, 29(7):  775-777. 

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Since the hospital information management has been constructed and applied in many hospitals and in most case the running of the system has been focused on technology and equipment,this paper discussed some important issues on the service capability that system offers.The key issue is how to use the information management system with two functions that provide information support to medical care and to help patients receiving the health care and medication.