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Table of Content

      
    10 February 2011
    Volume 49 Issue 2
    Articles
    Articles
    Relationship between serum 25-hydroxyvitamin D and the quality of life in middle-aged and older people
    WANG Yan, KANG Dong-hong, CAO Wei, WANG Ping, LIU Zhi-wen
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  1. 
    Abstract ( 1167 )   PDF (383KB) ( 186 )   Save
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    Objective    To assess the correlation of serum 25-hydroxyvitamin D (25OHD) and the quality of life in middle-aged and older people. Methods    A total of 167 middle-aged and older people, from 45 to 85 years old with a mean age of 61.02±10.06 in Jinan institutions, were recruited from September to November 2009. Serum 25OHD was categorized into 3 groups: <49.9nmol/L as the insufficient group, 50-74.9nmol/L as the borderline group, and ≥75nmol/L as the normal group. Serum 25OHD levels were measured by ELISA and the SF36 form was adopted to investigate the quality of life. Results    The mean 25OHD concentration was (54.21±19.88)nmol/L, while Vitamin D was detected as normal in 18.0% of the population. The 25OHD  concentration of females and males were (58.13±19.07)nmol/L and (51.24±20.07) nmol/L， respectively. The difference was significant (P<0.05). The 25OHD concentration of people below and above 60 yrs old were (58.82±19.34) and (49.54±19.43) nmol/L， respectively. The difference was also significant(P<0.01). People with insufficient vitamin D received lower scores than the other two groups in seven dimensions: physical functioning (PF), role-physical (RP), bodily pain (BP), general health (GH), vitality （VT）, social functioning （SF）, and mental health (MH) (P<0.05). Positive correlation between Vitamin D and the scores ofthe quality of life was found by Pearson analysis(P<0.05). Stepwise multiple linear regression showed that vitamin D was the main factor affecting the quality of life. Conclusion    25OHD insufficiency is prevalent in middleaged and older people, especially in the elderly and females. Lower serum 25OHD concentration in middle-aged and older people are associated with poorer quality of life. More attention should be paid to vitamin D supplementation in these populations.

    Role of Src kinase and NR2B in D1/D5 receptor agonist-induced spinal LTP
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  5-8. 
    Abstract ( 1017 )   PDF (718KB) ( 294 )   Save
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    Objective    To explore the role of Src kinase in the induction of spinal long-term potentiation (LTP) induced by the D1/D5 receptor agonist. Methods    C-fiber evoked field potentials were recorded at the superficial layer of the spinal dorsal horn at the lumbar enlargement by extracellular recording technique.  Results    ① SKF38393 (SKF, 250μmol/L)-induced LTP of C-fiber evoked field potentials was blocked by SCH 23390 (SCH, a D1/D5 receptor antagonist, 20μmol/L).  ② Genistein (200μmol/L), a selective Src kinase inhibitor, blocked the induction of spinal LTP induced by SKF.  ③ NR2B antagonist Ro256981 (Ro, 50μmol/L) prevented spinal LTP by SKF.  Conclusion    Activation of Src kinase and NR2B in the spinal dorsal horn may be crucial for the induction of spinal LTP by D1/D5 receptor agonists.

    Induction of apoptosis and G2/M cell cycle arrest by oridonin on human pancreatic cancer PANC-1 cells
    QI Xiao-li1, TIAN Ke-li1, ZHANG Dian-rui2, XU Xia1, FENG Fei-fei2, REN Gui-jie1, YUAN Hui-qing1, CHU Qian-qian1, ZHANG Qiang3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  9-13. 
    Abstract ( 1106 )   PDF (1610KB) ( 290 )   Save
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    Objective    To investigate the mechanism of oridonin on proliferation and apoptosis of PANC1 cells in vitro. Methods    The antiproliferation effect of oridonin was measured by MTT assay, PI staining and Hoechst 33342 staining were used to detect morphologic changes, the cell cycle distribution and the percentage of apoptosis were determined by flow cytometer, Caspase-3 activity was measured by spectrophotometry, and expression of pro-caspase-3, Bax and Bcl-2 were detected by Western blotting analysis. Results    Oridonin inhibited the growth of PANC-1 cells in a dose-dependent manner. Morphological changes indicated that oridonin induced PANC-1 cells apoptosis, the cell cycle was arrested at G2/M phase and the apoptosis rate increased in a dose-dependent manner, Pro-caspase-3 was activated, and expression of Bax was up-regulated, whereas expression of Bcl-2 had no obvious change. Conclusion    Oridonin can inhibit the growth of PANC-1 cells possibly by inducing G2 /M phase cell cycle arrest and apoptosis. The molecular mechanism of apoptosis may be mediated by the change of the Bax/Bcl-2 ratio and the activation of pro-caspase-3.

    Expressions of NMDA receptor subunits NR2B and NR2A and their relationship with apoptosis in the hippocampus of Alzheimer′s diseaselike rats
    LIU Zhi-an1, 2, ZHAO Wei-wei3, L Chun-e2, LI Mei2, XU Tie-jun1, 2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  14-18. 
    Abstract ( 1156 )   PDF (1279KB) ( 480 )   Save
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    Objective    To explore expressions of N-methyl-D-aspartate receptor subunits 2B (NR2B) and 2A (NR2A) and their relationship with apoptosis in the hippocampus of Alzheimer′s disease (AD)-like rats. Methods    The AD-like rat model was established by intracerebroventricularly injecting Aβ1-40 and intraperitoneally injecting AlCl3. Expressions of NR2B and NR2A in the AD-like rat hippocampus were detected by immunohistochemistry, and the apoptosis in the AD-like rat hippocampus was detected by TUNEL staining. Then the relationship was further explored between them. Results    Expression of NR2B was significantly increased in the AD-like rat hippocampus(P﹤0.05), especially in the CA1 subfield. Expression of NR2A had no significant change in the AD-like rat hippocampus(P>0.05). Apoptotic cells were significantly increased in the AD-like rat hippocampus (P﹤0.05), especially in the CA1 subfield, followed by the dentate gyrus and the CA3 subfield. Conclusion    NR2B, not NR2A, has pathological high expression in the AD-like rat hippocampus. This pathological high expression may have a specific relationship with AD-like apoptosis and the selective vulnerable phenomenon of the rat hippocampus.

    Effects of diazoxide on neuronal ultrastructure and free radicals in the  hippocampus of epileptic rats induced by lithium-pilocarpine
    SUN Xi-bo, HAN Yu-xiang, HAN Tao, WANG Xue, ZHAO Xiu-he, LIU Xue-wu, CHI Zhao-fu
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  19-23. 
    Abstract ( 993 )   PDF (990KB) ( 451 )   Save
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    Objective    To observe the effect of diazoxide(DZ), a mitochondrial ATPsensitive potassium channel (mitoKATP) opener, on neuronal ultrastructure and free radicals in the hippocampus of epileptic rats induced by lithium-pilocarpine, and explore the mechanism of the neuroprotective effect of DZ on epilepsy. Methods    Adult male Wistar rats were randomly divided into four groups: the control group, the epilepsy group (PILO group), the diazoxide group (DZ group), and the DZ and 5-hydroxydecanoate(5-HD) group (DZ+5-HD group). The model of status epilepticus (SE) was induced by lithium-pilocarpine. Before preparation of the model, the DZ group was treated with 5mg/kg of DZ, intraperitoneally injected (ip); and the DZ+5-HD group was treated with 8mg/kg of 5-HD, ip, then 5mg/kg of DZ, ip. The change of behavior in rats was observed. 4, 24 and 48 hours after SE, the rats were sacrificed and the hippocampi were removed. The neuronal ultrastructure was observed by an electron microscope. The content of malondialdehyde (MDA), and activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were measured with commercial assay kits. Results    Compared with the control group, hippocampal neurons in the PILO group and the DZ+5-HD group were severely damaged after SE. The number of neurons was decreased, and the neurons were swellen. There were numerous clumps distributed throughout the nucleus. Mitochondrial swelling was accompanied by the disappearance of cristae and disruption of membrane integrity. In more severe cases, the mitochondria were vacuolar. The damage was most severe 48h after SE. In the DZ group, the epileptic paroxysm latent period was significantly prolonged, the damage of neurons was less severe, and the mortality rate of rats with SE was decreased. In the DZ group, the MDA content was significantly decreased while the activities of SOD and GSH-Px were significantly increased compared with those in the PILO group. The effect of DZ could be abolished by 5-HD. Conclusion    The mitoKATP opener DZ can decrease the level of mitochondrial free radicals, thereby improve the response of the anti-oxidant system, and reduce oxidative stress after SE, so it can reduce neuronal injury and play an important role in neuroprotection after SE.  

    Construction of a eukaryotic expression vector containing the  Kir6.2 gene and its expression in HEK293 cells
    REN Xiao-yan, TAN Shu-hui, MA Guo-zhao， DU Yi-feng
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  24-28. 
    Abstract ( 1030 )   PDF (748KB) ( 488 )   Save
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    Objective    To clone the Kir6.2 gene, construct its eukaryotic expression vector，  and obtain positive HEK293 cell clones stably expressing the Kir6.2 gene. Methods    Total RNA was extracted from the SD rat brain. Full-length cDNA encoding the Kir6.2 gene was obtained by RT-PCR and inserted into the T1-simple cloning vector. After the sequencing was confirmed , the gene was subcloned into pcDNA3 to construct the recombinant eukaryotic expression vector pcDNA3.0/Kir6.2. The recombinant plasmid was transfected into HEK293 cells by lipofectamin and positive cell clones were screened with G418. Expression of the Kir6.2 gene in the transfected cells was confirmed by RT-PCR and Western blot. Results    PCR and sequencing showed that the target gene was cloned into the recombinant vector. Overexpression of the Kir6.2 gene in the transfected HEK293 cells was identified by RT-PCR and Western blot. Conclusion    The eukaryotic expression plasmid containing the Kir6.2 gene was successfully constructed. Positive HEK293 cell clones stably expressing the Kir6.2 gene were obtained.

    Protective effect of curcumin on dopamineinduced apoptosis in PC12 cells
    WANG Xiang-Yu1，MEI Yuan-Wu2，YANG Hong-Wei3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  29-33. 
    Abstract ( 1060 )   PDF (1313KB) ( 302 )   Save
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    Objective    To explore the protective effect of curcumin (Cur) on apoptosis of PC12 cells induced by dopamine (DA) oxidative stress. Methods    Using DA to induce PC12 cell damage, the model of oxidative stress-induced neuron damage was established. Proliferation of PC12 cells was observed by 3-［4 ,5dimethylthiazolyl ］-2 ,5diphenyltetrazolium bromide (MTT) assay. Morphology of the nucleus was observed by Hoechst staining. Apoptosis of PC12 cells was detected by propidium iodide(PI) staining flow cytometry (FCM). The mitochondrial membrane potential(△Ψm) was determined by Rh123 staining flow cytometry. Results    ① DA damaged PC12 cells in a concentration-dependent manner after application of DA (50-800μmol/L) for 24h, and MTT assay showed that Cur （10-40μmol/L） had no toxic effect on PC12 cells . ② Cur reduced the toxic effect induced by DA on PC12 cells in a concentration-dependent manner.  ③ Cur reducing the DA-induced toxic effect on PC12 cells was confirmed by cellular morphologic experiments. ④ PI staining FCM showed that Cur inhibited cellular apoptosis induced by DA and this effect was also confirmed by Hoechst staining. ⑤ The level of △Ψm was significantly decreased in PC12 cells after 100 and 200μmol/L DA treatment for 24h, which was significantly ameliorated by Cur (20 and 40μmol/L) treatment. Conclusion     Cur has a protective effect on PC12 cell damage induced by DA oxidative stress and this effect may be ascribed to the amelioration of the decreasing  level of △Ψm.

    Effect of advanced glycation end products on interleukin-1β and tumor  necrosis factor α secretion from microglial cells
    WANG Mei-xia, LIU Xue-ping, XU Song, DONG Chuan-fang, HOU Liang, YUAN Shu-hua
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  34-38. 
    Abstract ( 1083 )   PDF (779KB) ( 478 )   Save
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    Objective    To research the effect of advanced glycation end products (AGEs) on the levels of interleukin 1β(IL-1β) and tumor necrosis factor α(TNF-α)in primary rat microglial cells, and to further explore the effect of AGEs-BSA on Alzheimer′s disease(AD) and the possible mechanism at the cell ular level. Methods     Cultured microglial cells were intervened by AGEs-BSA and then identified with the immunocytochemistry method, and morphological changes of the cells were observed. After primary rat microglial cells were treated with 300μg/mL of AGEs-BSA and the RAGE neutralizing antibody, the levels of IL-1β and TNF-α extracted from the supernatant liquid of microglia were measured by enzyme-linked immunosobent assay(ELISA). Results     After the intervention of AGEsBSA, the cell body became bigger and the shape showed as an “Ameba”, and the levels of IL-1β and TNF-α were significantly increased（P＜0.001）. Compared with the AGEsBSA group, the levels of IL-1β and TNF-α were lower in cells exposed to the RAGE neutralizing antibody before treatment with AGEs-BSA （P＜0.01）, while they were higher than those in the normal control group（P＜0.01）. Conclusion      AGEs-BSA could activate microglia and induce the release of IL-1β and TNF-α in a time-dependent manner, which suggested that AGEs act directly or through the receptor activated microglia-mediated immune inflammatory responses.

    Proteomic study of differentially expressed proteins in the hippocampus of the rat model with  temporal lobe epilepsy
    RONG Jia1, CUI Ya-zhou2, ZHOU Xiao-yan2, SHANG Wei1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  39. 
    Abstract ( 1009 )   PDF (886KB) ( 542 )   Save
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    Objective     To study differentially expressed proteins in the hippocampus of rats with temporal lobe epilepsy(TLE) for exploring the pathogenesis mechanism and new therapeutic targets of epilepsy. Methods    Differentially expressed proteins in the hippocampus of rats with TLE induced by lithium-pilocarpine were analyzed and identified by using two-dimensional fluorescence difference gel electrophoresis(2D-DIGE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS).  Results     19 differentially expressed protein spots were detected in the hippocampus of rats with TLE compared with the control group, in which 13 were up-regulated and 6 were down-regulated. Conclusion     Many differentially expressed proteins exist in the hippocampus of rats with TLE, which may be related to the pathogenesis of epilepsy.

    Protective effects of SB202190 on hippocampal nerve cells in rats with  kainic acid-induced intractable epilepsy
    LIU Xue-wen1, ZHANG Hai-qi2, TIAN Bu-xian1, CAI Ai-min1, HAN Kun1, ZHANG Xue-jie1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  44. 
    Abstract ( 1055 )   PDF (1670KB) ( 538 )   Save
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    Objective    To explore the protective effect of p38 mitogen-activated protein kinase (MAPK) selective inhibitor SB20219 on hippocampal neurons in rats with kainic acid(KA)induced intractable epilepsy. Methods    50 SD rats were randomly divided into the negative group, the control group and the SB202190 treatment group, which was further divided into 3 subgroups，  the low dose group, the medium dose group and the high dose group. Behaviors were observed according to the criteria of Racine. Expressions of C-myc and Mkk3 were detected by immunohistochemistry and Western blot. The number of apoptosis neurons was observed and the apoptosis rate was calculated by TUNEL. Results    Behavior observation according to the Racine grading standard showed seizurefree in the negative group; the degree of attack was III-IV in the control group, while mostly I-II in the SB202190 treatment groups. Expressions of C-myc and Mkk3 were significantly decreased, and apoptosis cells reduced in the SB202190 treatment groups than those in the control group (P<0.01). Conclusion    SB202190 has protective effect on neurons of KA-induced intractable epilepsy rats. The mechanism may be associated with depressions of C-myc and Mkk3 expressions.

    Effects of early intensive insulin therapy on the dynamics of  blood glucose and immunoloregulation on T lymphocyte  subgroups in non-obese diabetic mice
    ZHANG Li-juan，LIU Cai-hong, HU Yan-yan, LI Gui-mei
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  49-53. 
    Abstract ( 1033 )   PDF (720KB) ( 425 )   Save
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    Objective    To investigate the immunoloregulation effects of early intensive insulin therapy on T lymphocyte subgroups in non-obese diabetic (NOD) mice. Methods    Thirty (12-14 weeks old)female NOD mice were randomly divided into the early intensive therapy(EIT) group, the early conventional therapy(ECT) group, the late intensive therapy(LIT) group, the late conventional therapy(LCT) group, and the no-therapy (DM)group. Also, age-matched female NOD mice, without the advent of diabetes, were included in the study as the normal control (NC) group. The changes of weight and blood glucose in the EIT group and other groups were observed and the ratio of T lymphocytes of spleen and thymus was analyzed by flow cytometer. Results    The weight of the EIT group increased with age compared with that in the LIT,  LCT and DM groups (P<0.001) and the average 24-hour blood glucose was sustained at (5.758±1.515) mmol/L, which was significantly lower than that in the LIT, LCT and DM groups(P<0.001) but was higher than that in the NC group. As for the T cells from the spleen, the percentage in the EIT group of CD4+ and CD3+ was low, and CD4+/CD8+ ratio significantly decreased compared with those of LCT and DM groups(all P<0.001). Compared with LIT, LCT and DM groups, the percentage of CD4+CD8- T cells from thymus in the EIT group was significantly lower(all P<0.001), while CD4+CD8+ was markedly higher(all P<0.001). Conclusions    Early intensive insulin therapy can effectively control blood glucose, keep normal weight gain, and decrease the host autoimmunity, which relate to the changes of T lymphocytes subsets.

    Coronary perfusion pressure during ventricular fibrillation and waveform-based  indicators for the return of spontaneous circulation
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  54. 
    Abstract ( 1244 )   PDF (1021KB) ( 369 )   Save
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    Objective   To evaluate whether coronary perfusion pressure （CPP） during untreated ventricular fibrillation (VF) and the corresponding VF waveform could be used to predict the probability of return of spontaneous circulation (ROSC).  Methods    VF was electrically induced and sustained for 8 minutes followed by 2 minutes of cardiopulmonary resuscitation and defibrillation in 20 pigs. Hemodynamics, VF mean frequency (VFmf) and VF mean amplitude (VFma) during untreated VF were analyzed by a multipurpose polygraph. According to the outcomes, the animals were categorized into the ROSC or the noROSC group. Results    There were 7 animals in ROSC group and 13 in no-ROSC group. Among ROSC animals, hemodynamic variables were better and both VFmf and VFma were higher than those of the no-ROSC animals (P<0.05). CPP, VFmf and VFma during untreated VF could be used to predict the probability of ROSC (area of ROC curve: 0.86, 0.76, and 0.82, respectively). Both VFmf  and VFma showed  positive correlations with CPP (r=0.67, r=0.71, respectively; P<0.01). Conclusions    Both CPP and VF waveform during untreated VF are valuable indicators for ROSC.

    Effect of  the β1-adrenoceptor antibody on potassium channels in  mouse cardiac myocytes
    SONG Ling-gang1, LI Xiao-dong2, MA Zhi-yong1, WANG Yuan-yuan1, WANG Jian-chun2, LI Li1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  58. 
    Abstract ( 1140 )   PDF (860KB) ( 266 )   Save
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    Objective    To investigate the influences of the adrenoceptor β1 antibody (anti-ADRβ1, simulating autoantibody) on potassium channels. Methods    Mouse ventricular cardiac myocytes were prepared by enzymatic isolating technique. The whole-cell patch-clamp technique was used to record K+ currents, in which signals were amplified with an HEKA EPC-10 patch clamp amplifier and controlled by Pulse software. The voltage protocol was a one-second depolarizing pulse from -50 to +50mV in 10mV increments and at a holding potential of -60mV. Cells were perfused with normal bath solution for 10 minutes to stabilize the currents, and then were perfused with anti-ADRβ1 in different concentrations (1/500, 1/100 or 1/50) for 5 minutes. Changes of transient outward potassium channel (Ito) and steady state potassium channel (Iss) current intensity and corresponding current-voltage relationships were monitored. Results    After perfusing cardiac myocytes with anti-ADRβ1 in different concentrations for 5 minutes, current intensity and corresponding current-voltage relationships of Ito did not significantly chang (P>0.05).  Compared with current intensity of Iss (4.7±0.24) pA/pF at normal bath solution, Iss (3.6±0.18) pA/pF  decreased in 1/500 concentration of antiADRβ1 but without statistical significance, however, Iss  significantly decreased in 1/100 (3.5±0.18) pA/pF and 1/50 (3.1±0.15)pA/pF concentrations of anti-ADRβ1 in an obvious dose-dependent manner.  Conclusion    The aadrenoceptor β1 Antibody inhibits Iss potassium channels of mouse cardiac myocytes but not Ito, which may play a role by stimulating β1-adrenergic receptors. The results provide experimental basis for further investigation of the adrenoceptor β1 autoantibody in heart failure.

    The model of atherosclerosis caused by different shear  stresses in the rabbit carotid artery 
    SUN Tao1, WANG Ying1, MA Zhi-yong1, ZHANG Kai1, ZHONG Xue-zhen2, LI Li1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  62-66. 
    Abstract ( 1045 )   PDF (744KB) ( 374 )   Save
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    Objective    To explore the effects of increased, lowered and oscillatory wall shear stresses in the progress of atherosclerosis by placing the arterial cannula in the rabbit carotid artery. Method    According to the theory of hydromechanics, the rabbit model of atherosclerosis was established by the arterial cannula designed by us and a high fat diet. The data was detected at three designated time points, including the common carotid arterial intimamedia thickness(IMT), end diastolic diameter(Dr), systolic peak velocity(Vs), diastolic end velocity(Vd), mean velocity(Vm). Then, the shear stress was calculated.  At the end of 12 weeks, the rabbits were sacrificed, and arterial samples were obtained to observe the pathological changes in common carotid artery through hematoxylin and eosin stain. Results    The levels of the increased and lowered shear stresses caused by the cannula were significantly different from the basic line(both P<0.05),  the vascular ultrasound showed there was turbulent blood flow in the area of oscillatory shear stresses. Compared to the  oscillatory shear stresses, the area of the lowered shear stress contained bigger plaques, more lipids and smooth muscle cells, but less collagen. There was no obvious plague in the area of the increased shearstress. Conclusions    The cannula in the common carotid artery can induce different shear stresses, including increased, lowered and oscillatory, in different segments of a single artery. Increased shear stresses can inhibit atherosclerosis, while the lowered and oscillatory ones can boost its formation.

    Effect of traditional Chinese medicine for kidney-reinforcingof morphological alterations of the adrenal gland in  mice with Kidney-Yang deficiency
    XU Cui-ping1，LI Zhen2，SONG Jie3，ZHANG Dan2，ZHU Qing-jun2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  67-70. 
    Abstract ( 1249 )   PDF (1492KB) ( 499 )   Save
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    Objective    To explore the effect of traditional Chinese medicine for kidneyreinforcing of  morphological alterations of the adrenal gland in mice with Kidney-Yang deficiency. Methods    The model of Kidney-Yang deficiency was established by overstrain and excessive sexual intercourse. The mice were randomly divided into three groups, including the normal control group, the Kidney-Yang deficiency group, and the Jinkuishenqi bolus group. After four-week treatment of traditional Chinese medicine for kidney-reinforcing, morphological changes of the adrenal gland were observed in all groups. Result    Compared with the normal control group, the weight and coefficient of the adrenal gland in the mice with Kidney-Yang deficiency were both obviously decreased（P﹤0.01; P﹤0.05）. Under a light microscope, it was shown that in the group of Kidney-Yang deficiency,   the adrenal cortex became thin, and the zona glomerulosa and fasciculate were narrow. The structure of cells was  irregularly arranged, while the cellular volume was reduced with shallow staining. In addition, vacuole-shaped cytoplasm and widened intercellular space were identified. Under an electronic microscope,  the shapes of nuclei were irregular in the glomerulosa zone of the adrenal cortex, while the nuclear membrane dissolved, the outer nuclear membrane protruded into the cytoplasm, and the chromatin were clumped. Compared with the normal control group, the number of mitochondria in the Kidney-Yang deficiency group was remarkably decreased, and parts of them were swollen and degenerated. The mitochondrial cristae were concentric or fingerprint-shaped and rough endoplasmic reticulum in the Kidney-Yang deficiency group were fewer than that in the normal control group. Smooth endoplasmic reticulum was obviously enlarged and vesicle-shaped. In contrast to the untreated group, the morphology of the adrenal cortex in the Jinkuishenqi bolus group was improved to a certain degree, which was similar to the normal control group. Conclusion     Morphology and ultra-structure of the adrenal gland in  mice with Kidney-Yang deficiency demonstrate degenerative changes. However, such damages can be repaired if treated with Jinkuishenqi bolus.

    Effects of extraneous adrenomedullin on Ang-Ⅱand MDA  expression in early renal trauma of rats
    WANG Xiao-mei1， ZHANG Shu-qin2， WANG Fang1， CHEN Chou-yan1， SUN Shao-hua1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  71-74. 
    Abstract ( 975 )   PDF (364KB) ( 419 )   Save
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    Objective    To investigate the effects of extraneous adrenomedullin(ADM) on Ang-Ⅱand MDA expression, and the protective action of ADM in early mechanical renal trauma.  Methods    104 healthy adult plain grade Wister rats were randomly divided into four groups: 8 in the control group, 32 in the trauma group,  32 in the group in which ADM was injected before trauma, and 32 in the group in which ADM was administered after the trauma. The model of mechanical kidney trauma was established by striking the skin area exactly above the kidney with a free falling ferrous hammer. In the latter two groups, ADM (0.1nmol/kg) was injected intraperitoneally 10 minutes before or after the striking. All rats were executed in batches at the 1st, 6th,12th, 24th hour after the trauma. The supernatant of renal homogenates was collected and the concentrations of Ang-II and MDA were detected by radioimmunoassay or colorimetry, respectively. Results     1) Concentration level of Ang-II: The concentration of Ang-II in the trauma group was significantly higher than that in the control group in the early trauma (1st, 6th hr) (P<0.05). Also, both the concentrations of Ang-II in the group injected with ADM before trauma (1st, 6th hr) and the group injected  with ADM 1hr after trauma at were lower than that in the trauma group (P<0.05). 2) Concentration level of MDA: The concentration of MDA in the trauma group was obviously higher than that in the control group at 1 h (P<0.05). But the concentrations of MDA either in the group injected with ADM 6 hrs before trauma or the group injected with ADM 12 hrs after trauma were lower than that in the trauma group (P<0.05). Conclusions     Extraneous ADM can exert a protective effect onrenal trauma by controlling the concentration of Ang-Ⅱand MDA. More importantly, ADM injected before trauma shows stronger effect on renal trauma than that injected after trauma.

    Effects of high-concentration distilled water and cisplatin on  osteosarcoma cells in a local and shorttime manner
    QI Lei, LI Jian-min, XIANG Long-zhan, YANG Qiang
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  75-78. 
    Abstract ( 1159 )   PDF (1184KB) ( 470 )   Save
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    Objective    To study the effects of high-concentration distilled water(DW) and cisplatin administered locally in a short time on osteosarcoma (OS) cells. Methods    Cultured osteosarcoma cells, U2-OS, were divided into the DW group, the DW31.25 group, the DW62.5 group, the DW125 group, the DW250 group and the DW500 group according to the corresponding concentrations of cisplatin applied, i.e. 31.25μg/mL, 62.5μg/mL, 125μg/mL, 250μg/mL and 500μg/mL, respectively. Then, osteosarcoma cells were processed for 5 minutes. At 0, 24, 48, 72 and 96h, the inhibition rates of tumor cells were determined by MTT assay. Changes of cells in morphology and quantity were observed under inverted microscopy. Results    Among the groups, there was no significant difference in the inhibition rates at 0h. Though no differences were observed among the DW62.5 group, the DW125 group, the DW250 group and the DW500 group at 96h, the inhibition rates were significantly higher than those of the DW group and the DW31.25 group. In the former 4 groups, hardly any osteosarcoma cells survived at 96h while massive osteosarcoma cells survived in the DW group. Conclusion    Highconcentration distilled water and cisplatin administered in a local and short-time manner produces strong killing effect on osteosarcoma cells.

    Astragalus polysaccharides down-regulate expression of  α-SMA in airway smooth muscle cells
    ZHANG Xue-ping, WANG Li, LONG Fei, LI Huai-chen
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  79. 
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    Objective    To investigate the effect of astragalus polysaccharides (APS) on expression of α-smooth muscle actin(α-SMA) in airway smooth muscle cells, and to investigate the possible mechanism of the effect. Methods   APS were isolated from Astragalus membranaceus. Cultivated human airway smooth muscle cells were divided into the APS groups in doses of 20, 100 and 500μg/mL, the control group, the lipopolysaccharide (LPS) group and Toll-like receptor 4(TLR4) antagonist group, and subsequently stimulated to express α-SMA by TGF-β. Expressions of α-SMA and mRNA were measured by immunocytochemistry, Western blot and RT-PCR. Results    Medium and high concentrations of APS inhibited expression of αSMA(P<0.01) and down-regulated the level of α-SMA mRNA(P<0.01), which could be partly reversed by the TLR4 antagonist (P<0.01). Conclusion    APS down-regulates expression of α-SMA stimulated by TGF-β in airway smooth muscle cells through TLR4,and has a potential role in the prevention and treatment of asthma.

    Clinical characteristics and surgical treatment of pulmonary  sclerosing hemangioma
    FANG Jian-wei1, LIU Fan-ying1, LI Jun1,2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  83-87. 
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    Objective    To investigate the clinical characteristics, diagnosis and treatment of pulmonary sclerosing hemangioma (PSH) and to deepen the understanding of PSH. Methods    Clinical data from 18 patients with PSH, who underwent surgical resection in the Provincial Hospital Affiliated to Shandong University from April 2004 to March 2010, and the related literature were retrospectively reviewed. Results    Of the 18 patients with PSH confirmed by histopathology, 2 were males and 16 were females. The median age was 41.6 years, ranging from 18 to 56 years. 12 patients were detected by physical examination, while 4 patients presented with cough, 1 with cough and chest pain, and 1 with blood-stained sputum. The imaging examination generally revealed the peripherally solitary, well-circumscribed, and round or similarly round nodule with homogeneous density, and partly punctiform calcification. All these 18 patients received surgical resection without complications and mortality, 8 with lobectomy, 1 with wedge resection and 9  with tumor enucleation. No recurrence and no metastasis were detected during follow-ups. Conclusions    Clinical characteristics and imaging examinations of PSH are lack of specificity, and the accurate diagnosis depends on pathological examination. PSH should be suspected in middle-aged female patients with a peripherally solitary, well-circumscribed, and round or similarly round nodule of homogeneous density, whether the patients present symptoms or not. Surgical resection is effective for PSH.

    Inheritance structure of the human Tim-3 gene in the Uigur minority  population from Xinjiang Province
    WANG Yan, HU Hai-fang, ZHANG Hua
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  88-92. 
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    Objective    To detect single nucleotide polymorphisms(SNPs) in the promoter and non-translational region of the human T-cell immunoglobulin and mucin domain-3(Tim-3) gene in the Uigur of the Xinjiang Uygur Autonomous Region, and to search for genetic marks of the Tim-3 gene. Methods     Employing direct sequencing PCR amplification,  the lengths of 5′promoter region 3kb and 3′ext 1kb of the Tim-3 gene were detected to screen for SNPs in 32 common Uigurs of Xinjiang Province, and the results were compared with other ethnic groups in the database of the National Center for Biotechnology Information (NCBI). The position, type and frequency of the Tim-3 gene in Uigurs of Xinjiang Province were determined. Results    12 SNPs were detected in the promoter and non-translational region of the Tim-3 gene, including 8 existing high-frequency SNPs and 4 newly-discovered low-frequency SNPs. The genotype and allele distribution of each SNP proved to attain heredity balance according to the Hardy-Weinberg equilibrium law, possessing a groupspecialty. The distribution and frequency of the 32 SNPs showed no statistical difference. Conclusion    The SNP distribution of the Tim-3 gene in Uigurs of Xinjiang Province has racial and regional differences, which may lay the foundation for the study about the relativity between the Tim-3 gene and diseases in the Uigur and Han.

    Effect of Atorvastatin combined with Amiodarone on prevention of paroxysmal atrial fibrillation after dual-chamber-pacemaker implantation
    WEI Zi-xiu, ZHANG Mei
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  93-96. 
    Abstract ( 1380 )   PDF (361KB) ( 360 )   Save
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    Objective    To evaluate the effect of atorvastatin combined with amiodarone on preventing paroxysmal atrial fibrillation(PAF) following dual-chamber-pacemaker implantation in the patients with sick sinus syndrome(SSS)．  Methods     Ninety-four SSS patients suffering from PAF after dual-chamber-pacemaker implantation were randomly divided into the treatment group and the control group． While the patients in the control group were given amiodarone only,  those in the treatment group were administrated with additional atorvastatin (20mg/day). After a 12-month followup, PAF times, AF duration and serum level of C-reactive protein(CRP) were observed． Results     PAF times, AF burden，  AHREs recorded by the pacemaker, the effectiveness of AF treatment and serum level of CRP in the treatment group were significantly lower than in the control group(P<0.05).  Conclusions     Atorvastatin combined with amiodarone can effectively inhibit the occurrence of PAF after dual-chamber-pacemaker implantation in patients with SSS． The mechanism may be attributed to the effect of atorvastatin to inhibit Inflammatory reaction．

    Cell cycle distributions and cell proliferation-related protein expression in  myelodysplastic syndromes, aplastic anemia, and acute leukemia
    WANG Rui-jie1, XU Hong-zhi1, HUANG Min3, MA Chun-yan2, SUI Xiao-hui1, LIU Xin1, ZHANG Bing-chang3, LI Yuan-tang3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  97-101. 
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    Objective    To explore cell cycle distributions and expressions of CD34 and Ki67, and cell proliferation-related proteins in bone marrow mononuclear cells(BMMNCs) among myelodysplastic syndromes(MDS), aplastic anemia(AA), and acute leukemia(AL) patients. Methods     Bone marrow aspirates were collected from 68 patients between June 2009 and June 2010 at the Department of Hematology, Provincial Hospital affiliated to Shandong University.The cases consisted of 30 MDS (18 refractory anemia and 12 refractory anemia with excess blasts), 22 AA (7 severe  and 15 non-severe cases), and 18 AL (12 acute myeloid leukemia and 4 acute lymphoblastic leukemia). 18 healthy individuals without any hematological problems were used as normal controls. Propidium iodide and immunofluorescent double staining through flow cytometry were applied to explore the cell cycle distributions and expression of CD34 and Ki67. Results     Compared with the control group, the percentages of G0/G1 phase of the cell cycle, haemopoietic stem/progenitor cells CD34+ cells, Ki67+ cells, CD34+Ki67+ cells, Ki67+ cells in CD34+ cells, and CD34+ cells in Ki67+ cells of BMMNCs were all significantly increased in the MDS and AL group(P<0.05), whereas the percentages of S and S+G2/M phases of the cell cycle were both significantly decreased(P<0.05). In addition, the proportion of G2/M phase of the cell cycle was lower in the AL group than the control group(P<0.05). The ratios of G0/G1 phase, CD34+ cells, Ki67+ cells, CD34+Ki67+ cells, Ki67+ cells in CD34+ cells, and CD34+ cells in Ki67+ cells were significantly higher in RAEB(refractory anemia with excess blasts) compared to RA(refractory anemia) and in AL compared to RAEB  (P<0.05), accompanied by lower ratios of S and S+G2/M phases (P<0.05). There were no significant differences in the ratios of G0/G1, S, G2/M, and S+G2/M phases of BMMNCs between the AA and the control group (P>0.05), whereas the proportions of CD34+, CD34+Ki67+, Ki67+ cells in CD34+ and CD34+ cells in Ki67+ cells were all significantly lowered in the AA group (P<0.05). The percentages of G0/G1 phase, CD34+ cells, Ki67+ cells, CD34+Ki67+ cells, Ki67+ cells in CD34+ cells, and CD34+ cells in Ki67+ cells were  more significantly increased in the RA group than in the AA group (P<0.05), whereas the ratios of S and S+G2/M phases were both significantly decreased (P<0.05). Conclusion     This study suggests that AA is different from MDS and AL. There is similar pathogenesis for MDS and AL. Analyses of the cell cycle and cell proliferation in bone marrow mononuclear cells are reliable approaches for the diagnosis and differential diagnosis of RA and AA.

    Comparison of morphine and fentanyl on systemic inflammatory reaction and myocardial function in cardio-valve  replacement surgery under CPB
    MENG Tao1, ZHANG Xin-yu2, LI Xing-hua2, YU Jin-gui1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  102-105. 
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    Objective    To compare the effects of morphine and fentanyl on systemic inflammatory reaction and myocardial function in patients undergoing cardiovalve replacement under cardiopulmonary bypass(CPB).  Methods    Thirty patients (ASAⅡ-Ⅲ) undergoing cardio-valve replacement surgery under CPB were randomly divided into two groups (n=15). Anesthesia was induced with intravenous injection of midazolam, morphine (group M) or fentanyl (group F) and subsequently pipecuronium, and was maintained with interval injection of morphine (group M) or fentanyl (group F) and pipecuronium. Serum SOD, MDA, TNF-α, IL-6, IL-8, IL-10, CK-MB and cTnI was detected before anesthesia (T0), before skin incision (T1), before aortic-clamping (T2), 2h(T3) and 6h (T4) after CPB and 24hafter surgery (T5). Results    Serum levels of SOD activity, MDA, TNF-α, IL-6, IL-8, IL-10,  CK-MB activity andcTnI increased at different time points of post-CPB (P<0.05-0.01), and the extent, except for serum SOD activity, in group M was lower than that in group F (P<0.05). There were no differences in cardiac recovery and usage of cardiovascular activators between the two groups. Conclusion    The findings of the present study demonstrate that morphine can inhibit the release of oxygen free radicals and the inflammatory response, and protect the myocardium more than fentanyl in patients undergoing cardiovalve replacement under CPB.

    Expressions of eIF4E and MMP-9 in lesions of psoriasis vulgaris
    CAI Da-xing1, MA Wei-yuan1,2, SUN Qing1, ZHONG Hua1, LI Chun-yang1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  106. 
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    Objective    To study expressions and roles of eukaryotic translation initiation factor 4E (eIF4E) and matrix metalloproteinase-9 (MMP-9) in lesions of psoriasis vulgaris (PS). Methods    Samples were collected from 40 PS patients and 30 healthy controls under informed consent. Expressions of eIF4E and MMP-9 in the samples were detected with the PV-9000 immunohistochemical method. Results    ① No expression of eIF4E was observed in the normal samples, while weak positive and positive expressions were observed in the lesions of PS. ② MMP-9 was expressed in both samples, and the levels of eIF4E and MMP-9 were significantly higher in the lesions of PS than those in the normal skin tissue(P<0.01).  ③ There was no correlation between expressions of eIF4E and MMP-9 in psoriatic lesions. Conclusion    Both eIF4E and MMP-9 are overexpressed in the lesions of PS, and may contribute to the pathogenesis of psoriasis.

    Expression of HGF, C-Met, VEGF-A and their correlations with  malignancy and angiogenesis in astrocyte tumors
    LIU Bin, LIU Wei, FAN Hai-tao, HOU Xian-zeng, JIA Jian-bo, XU Shang-chen, PANG Qi
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  109. 
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    Objective    To explore expression of VEGF(vascular endothelial growth factor), HGF(hepatocyte growth factor) and C-Met, the receptor of HGF, in low- and high-grade astrocytoma and their correlations with angiogenesis, pathological differentiation and prognosis. Methods    Protein expression of HGF, C-Met and VEGF was examined with the immunohistochemical staining Method (S-P). mRNA expression of HGF, C-Met and VEGF was examined with RT-PCR. Results    Expression rates of HGF, CMet and VEGF in 98 astrocyte tumors were 98.81%, 97.82% and 100%, respectively, and the degree of expression was in accordance with malignancy of the tumors. The MVD of astrocyte tumors was positively correlated with the malignancy. Such correlations existed among HGF, C-Met and VEGF. Conclusion    Up-regulated expression of HGF, C-Met and VEGF are inseparably and closely related to malignancy and angiogenesis of the tumor, and expression of protein and mRNA can act as an indicator of the prognosis for the patients with astrocyte tumors.

    Clinical effectiveness of dendritic cells therapy combined with  intravesical  perfusion of epirubicin on bladder cancer after TURBT
    NI Yong-liang, LI Qing
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  114-118. 
    Abstract ( 1094 )   PDF (459KB) ( 205 )   Save
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    Objective    To evaluate the effectiveness of intra-dermal injection of dendritic cells combined with intra-vesical perfusion of epirubicin (EPI) on preventing recurrence of bladder cancer after TURBT. Methods    106 bladder cancer patients were randomly divided into two groups after TURBT， group A was treated with intradermal injection of dendritic cells combined with intravesical perfusion of EPI, while group B was treated only with intra-vesical perfusion of EPI. All patients were followed up for at least 2 years. T-lymphocyte subsets,tumor recurrence rates and adverse reactions of patients were observed and compared.  Results    CD3+, CD4+ and CD4+／CD8+ of group A increased after dendritic cells therapy. Compared with that before the therapy and that of group B, the differences were significant(P<0.05). 2-year recurrence rates of group A and group B were 10% and 25.8%, respectively. The difference was significant(P<0.05). 3 patients of group A developed modent dermal inflammation after  dendritic cells therapy. Conclusions    Compared with simple intra-vesical perfusion of EPI , DC therapy combined with intravesical perfusion of EPI can effectively reduce 2-year recurrence rate of bladder cancer after TURBT.Therefore, DC immune therapy combined with intra-vesical chemotherapy provides a new way for the adjuvant treatment of bladder cancer.

    Application of the geographically weighted regression model in  spatial genetic structure of the human population
    LI Xiao, XUE Fu-zhong
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  119-124. 
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    Objective    To explore the application of the geographically weighted regression(GWR) model in analyzing influencing factors of human population genetic structure. Methods    Using global ACE gene data and climate surveillance data, based on latent variable analysis and spatial statistical methods, synthesis factors from the climate variables were extracted by confirmatory factor analysis. Then spatial distribution of the ACE gene D allele frequency and the climate synthesis factors were estimated with the Kriging interpolation method. Finally the multiple linear regression model (global model) and GWR model (local model) were constructed to explore the relationship between the D allele frequency and the climate synthesis factors, respectively. Results     Two latent synthesis factors were extracted by confirmatory factor analysis. The multiple linear regression model showed that the two synthesis factors were both statistically related with D allele frequency (P<0.01). The local R2 and parameter estimation of each spatial unit of the GWR model displayed significant spatial variability, and its fitting effect was more desirable compared with the global model. Conclusion    The GWR model is more accurate in describing the spatial varying relationship between human population genetic structure and climate factors, and it is obviously superior to the global model.

    Management and flow direction of multi-drug resistant tuberculosis  patients discharged from TB hospitals
    ZHANG Jia-sheng1, SUN Qiang1, BIAN Xue-feng1,2, YAN Yun1, LI Wen-jing1, LI Ren-zhong3, CHEN Cheng3, ZHANG Hui3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  125. 
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    Objective    To investigate the management and flow direction of multi-drug resistant tuberculosis（TB） patients after discharged from the TB hospitals.  Methods    The samples were randomly selected using the stratified sampling method. Questionnaires were used in multidrug resistant tuberculosis patients in TB hospitals to get quantitative data and the information was further analyzed by SPSS 16.0. The semi-structured questionnaire was applied in the key subjects. Results    Among the patients， 7.2% were transferred to TB dispensaries after being discharged from TB hospitals, 8.3%  took medication under doctors′ supervision, and 35.1% received visits from personnel of TB dispensary. Conclusions    Only few patients are referred to TB dispensaries, and the management of multi-drug resistant tuberculosis patients is not adequate. So,  cooperation between TB hospitals and TB dispensaries needs to be strengthened in order to control multi-drug resistant tuberculosis.

    Epidemiological status and related risk factors of COPD in  urban residents in Jinan
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2011, 49(2):  128. 
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    Objective    To obtain the prevalence of chronic obstructive pulmonary disease (COPD) in urban residents in Jinan， to learn the status of diagnosis and treatment and to explore the possible related risk factors.   Methods    A cross-sectional study with a multistage stratified random sampling method was performed to select subjects aged ≥ 40 years. A questionnaire, designed with reference to the epidemiological questionnaire made by the program of COPD burden of GOLD committee and the practical situation of China, was used in interviews of the subjects. Then the CHEST HI-801 spirometer was used to test lung function in those who finished the questionnaire. The diagnosis of COPD was based on the standard made by the COPD group of Chinese Medical Association (2007 version). Results    According to the sampling rule, 2467 subjects should be included, among whom 2055 subjects finished the questionnaire and spirometry. The overall prevalence of COPD in urban residents in Jinan was 7.6%(156/2055), 12.2%(111/156) in males and3.9%(45/156) in females. 84 cases had been confirmed, in which only 37 (44.0％) cases accepted regular treatment,  and 48（30.8%） cases were asymptomatic. Multivariate logistic regression analysis found that the main factors affecting the state of COPD were smoking, males, senility, low income and a family history of respiratory diseases.  Conclusion    The overall prevalence of COPD in urban residents aged ≥ 40 years in Jinan was 7.6%. The actual state of diagnosis and treatment is far from satisfactory. Risk factors of COPD are related to individuals, family, and society.

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