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Table of Content

      
    10 February 2013
    Volume 51 Issue 2
    Articles
    Valsartan alters the balance of Th17 cells to regulatory T cells in chronic viral  myocarditis
    LU Guan-yan1, CUI Bin2, LIU Zhong-liang3, LI Yu-tang4, LIU Xue-fei3, MA Xiao-jing1, ZHU Gui-yue1, YUAN Hai-tao1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  1. 
    Abstract ( 843 )   PDF (3673KB) ( 337 )   Save
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    Objective   To observe  the effects of angiotensin receptorⅡ blockers(ARB) Valsartan on the balance between  Th17 and Treg cells in chronic viral myocarditis (VMC) and explore the effects of ARB on chronic VMC and its mechanisms. Methods   BALB/c mice(n=36) were infected with CVB3 at day 1,14 and 28 to establish the chronic VMC models.The volumes of CVB3 were 0.20,0.25 and 0.30mL.The mice in the normal control group(n=6) were intraperitoneally injected with equal volumes of  normal saline without CVB3 at the same time.The survival mice infecteded with CVB3 were randomly divided into the chronic  VMC control group and the Valsartan therapy group at day 42. The mice of the Valsartan therapy group were administered with Valsartan 10mg/(kg·d) for 28 days. Blood pressure, heart weight and body weight of all the mice were  measured before execution. Lymphatic cells were extracted from the spleen and the ratio of Th17 and Treg cells was evaluated by flow cytometry. mRNA of IL-17A and foxp3 were tested by Real-time PCR.The severity of myocarditis was assessed by haematoxylin-eosin(HE) and Masson’s trichrome. Echocardiography was performed to evaluate  cardiac functions of the mice. Results   There was no significant difference in  blood pressure level among the three groups(P>0.05). Compared with  Valsartan group and normal  control group, the ratio of heart weight in  chronic VMC group was higher,and the percentage of CD4+Th17 cell and the level of IL-17 in  chronic VMC  group were significantly up-regulated(P<0.01), while the pencentage of CD4+CD25+ Treg cell and cytokine of foxp3 was down-regulated (P<0.01, P<0.05 respectivcly). Cardiac functions of the mice in  Valsartan therapy group increased significantly. Conclusion   Th17/Treg imbalance exists in mice with chronic VMC. Valsartan can regulate Th17/Treg imbalance to ameliorate viral myocarditis and the progression of  dilated cardiomyopathy.

    Expression and location of Wnt7b in  rat subgranular zone of  hippocampus dentate gyrus after cerebral ischemia-reperfusion
    LI Hui1,HUANG Jing-yang1, CHEN Hai-li1, LIU Bao-yi2, YUAN Zhong-rui1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  7-11. 
    Abstract ( 880 )   PDF (3177KB) ( 272 )   Save
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    Objective   To investigate the involvement of Wnt in ischemia-induced adult nerve regeneration and explore the mechanism of the two members.Methods   44 normal male Wistar rats were randomly divided into 2 groups: the normal control group and the cerebral ischemia group. Cerebral ischemia-reperfusion rat models were established by inserting a proper thread in the right middle cerebral artery and removing the thread after 90min.Infarction and neurological deficit scores were evaluated after the operation. RT-PCR was used to investigate the expression of Wnt7b mRNA in hippocampus. Immunohistochemistry was used to investigate  expressions of Wnt7b and β-catenin proteins in the dentate gyrus of hippocampus. Results   Severe infarction appeared 1 day after ischemia-reperfusion, with  obvious neurological deficit. Neural function′s were  improved as time progressed. Expressions of Wnt7b mRNA in health side and  ischemic side were significantly upregulated, and  it  was more obvious in  ischemic side  (P<0.01)than  in contralateral side(P<0.05). Consistent with the level of Wnt7b mRNA, the amount of Wnt7b positive cells dramatically increased in close proximity to the subgranular zone (SGZ) of the dentate gyrus in hippocampus(P<0.01), and the immunostaining of β-catenin, a primary element of classical Wnt signalling pathway, also significantly increased(P<0.01). Conclusion   Ischemia injury stimulated the expressions of Wnt7b and β-catenin in SGZ of hippocampus in adult rats. Wnt7b might be involved in the neurogenesis in SGZ of adult hippocampus after  cerebral ischemia by the classical Wnt signalling pathway.

    Construction of a lentiviral vector carrying shRNA against HIF-1α and generation of a GL261 cell line stably transfected with this vector
    XU Yang-yang1,2, JIANG Zheng1,2, ZHOU Wei3, JIANG Yu-quan1, 2, LI Xin-gang1, 2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  12-16. 
    Abstract ( 966 )   PDF (9164KB) ( 342 )   Save
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    Objective   To construct a lentiviral vector carrying a short hairpin RNA (shRNA) targeting the HIF-1α gene and detect the silencing effect of the vector on mouse glioma cell line GL261. Methods   Four double-stranded shRNA targeting the HIF-1α gene were designed, synthesized and cloned. The resulting lentiviral vector containing HIF-1α shRNA was named as pLenti6.3-shRNA3. GL261 cells were transfected with pLenti6.3-shRNA3 lentivirus to obtain a cell line stably expressing HIF-1α shRNA. After the transfection, mRNA and protein expressions of HIF-1α in GL261 cells were detected by real-time PCR and Western blot, respectively. Results   A lentiviral vector carrying a shRNA targeting HIF-1α gene was successfully constructed. The GL261 cell line stably transfected with the vector was established. The recombinant lentivirus were harvested from 293T cells with titer of 1×108 TU/mL. Real-time PCR and Western blot analyses confirmed that the expressions of HIF-1α was down-regulated in GL261 cell line which stably transfected with the recombinant vector. Conclusion   shRNA targeting different sites of the HIF-1α gene exhibits different inhibitory effects. Specific shRNA could induce stable silencing of HIF-1α gene.

    Role of angiotensin Ⅱ inducing cell senescence in rat hippocampal neurons in primary culture
    ZHANG Na-na1, WANG Li-xiang1, ZENG Ji-ping2, WEI Xin-bing1, CAO Min-min1, LIU Hui-qing1, SUN Xia1, ZHANG Xiu-mei1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  17-21. 
    Abstract ( 886 )   PDF (53462KB) ( 336 )   Save
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    Objective   To investigate the effect and related mechanism of angiotensin Ⅱ (Ang Ⅱ) inducing cell senescence in rat hippocampal neurons in primary culture. Methods   Primary neonatal rat hippocampal neurons were cultured. Senescence-associated β-galactosidase (SA-β-gal) staining was used to detect  cell senescence induced by Ang Ⅱ in different concentrations and at different time to determine the best inducing condition. To further explore the mechanisms, the effects of AT1 receptor blocker irbesartan and AT2 receptor blocker PD123319 on Ang Ⅱ-induced cell senescence were examined. Expressions of cell cycle-related protein P53 and P21 were detected by Western blot assay. Results   Forty-eight his incubation of Ang Ⅱ (10μmol/L)  could induce cell senescence in  primary rat hippocampal neurons. Irbesartan could  inhibit cell senescence induced by Ang Ⅱ and reduce the expression of P53 and P21, whereas PD123319 had no effect on both of them. Conclusion   Ang Ⅱ can induce cell senescence in the primary rat hippocampal neurons. The effect of Ang Ⅱ on cell senescence may be mediated by AT1 receptor and the mechanism may be related to the over-expression of protein P53 and P21.

    Effects of eye acupuncture on VEGF and VEGFR expressions in focal cerebral ischemia-reperfusion rats
    MI Dan1,2, MA Xian-de3, WANG Jian4
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  22. 
    Abstract ( 840 )   PDF (3397KB) ( 405 )   Save
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    Objective   To explore the mechanism of eye acupuncture in promoting  microvessel generation of focal cerebral ischemia-reperfusion rats by observing its effects on expressions of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor (VEGFR). Methods   Improved suture-occluded method was adopted to establish focal cerebral ischemia-reperfusion model, and eye acupuncture was applied. Immunohistochemistry was employed to detect the microvessel density and expressions of VEGF and VEGFR2 in the brain after 3, 24 and 72h reperfusion. RT-PCR method was used to detect the expressions of VEGF mRNA and VEGFR2 mRNA. Results   Compared with model group, the microvessel density was significantly increased 24h and 72h after ischemia/reperfusion in the eye acupuncture group, the expressions of VEGF and VEGFR2 at each point were also markedly increased (P<0.05), and the expressions of VEGF mRNA and VEGFR2 mRNA at each point were conspicuously regulated up (all P<0.05). Conclusion   Eye acupuncture can promote the microvessel generation of cerebral ischemia-reperfusion rats, and the mechanism is related to up-regulated expressions of VEGF/VEGFR.

    Effects of PD98059 combined with cisplatin derivative on human ovarian carcinoma cell SKOV3
    JIA Zong-yang, ZHANG Xiang-ning, JIN Yan-hui, WEI Mei-ling
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  27-32. 
    Abstract ( 718 )   PDF (1932KB) ( 476 )   Save
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    Objective   To investigate the effects of MEK/ERK inhibitor PD98059, cisplatin and the combined treatment on the proliferation of human ovarian carcinoma cell SKOV3, and to investigate the potential mechanism. Methods   Cell Counting Kit-8(CCK-8)was used to investigate the SKOV3 cell proliferative inhibition rates of different groups: the blank control group, the cisplatin group, the PD98059 group and the combined treatment group. Real-Time PCR was used to detect the expression levels of WW domain-containing Oxidoreductase (WWOX) and extracellular regulated protein kinases (ERK) mRNA. Western blotting analysis was used to measure the expression of phosphorylated ERK1/2(p-ERK1/2) in different groups. The morphological changes of various groups were observed under the light microscope. Flow cytometry was employed to analyze the apoptosis rate of SKOV3 cells in different groups. Results   The inhibitory effect on the proliferation of SKOV3 cells in the combined treatment group was more significant compared with the other three groups (P<0.05). RT-PCR analysis showed that the expressions of WWOX mRNA were increased in the cisplatin group and the combined treatment group compared with the other two groups. The WB analysis showed that the expressions of p-ERK1/2 in the PD98059 group and the combined treatment group were higher compared with the other two groups. More adherent cells were observed in the blank control group and the PD98059 group under the light microscope. There were less adherent cells in the cisplatin group and the combined treatment group, and cytoplasmic dissolution of cells could be observed under the light microscope in these two groups. Conclusion   PD98059 combined with cisplatin exhibited significant inhibitory effect on the proliferation of SKOV3 cells via regulating the expression of WWOX gene and the ERK pathway.

    Establishment of a bortezomib-resistant cell line KM3/BTZ of human multiple myeloma
    PU Ye-di, LI Li-zhen, MA Dao-xin, DONG Ke,ZHAO Chuan-li, SONG Qiang, WANG Lu-qun
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  33. 
    Abstract ( 1264 )   PDF (1597KB) ( 470 )   Save
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    Objective   To establish a bortezomib-resistant cell line KM3/BTZ of human multiple myeloma (MM) and investigate its biological characteristics and mechanisms of drug-resistant in MM preliminarily. Methods   The KM3 cell line was exposured to bortezomib at increasing doses to obtain a stable bortezomib-resistant KM3/BTZ in vitro. The growth curve was drawn and the doubling time was counted. MTT assay was used to evaluate the sensitivity of drug resistance. The expression of MDR-1 mRNA was determined by RT-PCR in KM3 and KM3/BTZ cell lines. Results   The KM3/BTZ cell line resistant to bortezomib was established successfully with the resistance index of 19.7 (KM3/BTZ to KM3). Compared with the parent cells, KM3/BTZ exhibited a significant longer doubling time (P<0.05) .The expression of MDR-1 mRNA was not observed in either resistant or parent cells. Conclusion   We have successfully established bortezomib resistance cell line KM3/BTZ and its drug-resistant character is stable. The KM3/BTZ cell line might serve as an ideal model to explore the drug-resistance mechanisms and to reverse the drug resistance.

    Poly(ADP-ribose) polymerase 1 inhibition protects human bronchial endothelial cells against nicotineinduced inflammation response
    PENG Xiao-nu1,2, LI Wen-jun2, TIAN Hui1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  37-43. 
    Abstract ( 909 )   PDF (7390KB) ( 354 )   Save
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    Objective   To detect the changes of bronchial epithelial cells inflammatory cytokine stimulated by nicotine and to investigate the function and mechanism of poly(ADP-ribose) polymerase 1 (PARP1) in the inflammatory response mediated by toll-like receptor 4 (TLR4). Methods   Bronchial epithelial cells were cultivated. The expressions of inflammatory cytokine-related gene and protein were detected in nicotine stimulation group, TLR4 and PARP1 inhibition group and their negative control groups. Results   Compared with the controls, nicotine stimulation increased the protein expressions of TLR4 and PARP1. TLR4 inhibition reduced nicotine-induced up-regulations of iNOS, ICAM-1 and PARP1. NF-κB inhibition decreased ICAM-1 and iNOS expressions. PARP1 inhibition decreased protein expression of inflammatory cytokines induced by nicotine stimulation, probably through preventing NF-κB nuclear translocation. Conclusion   Nicotine increases ICAM-1 and iNOS expressions via TLR4/PARP1/NF-κB pathway. PARP1 might be a dispensable factor in TLR4-mediated inflammation after nicotine stimulation. PARP1 inhibition might shed a light on the decrease of nicotine-induced inflammatory cytokines expression during bronchial epithelium atypical heperplasia.

    Mechanisms involved in hydrogen sulfide attenuating high glucose-induced inflammation in endothelial cells
    WANG Xiao-lei1, GAO Ling1, YU Chun-xiao1, LIU Yuan-tao2, ZHANG Nan1, GUAN Qing-bo1, ZHAO Jia-jun1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  44-48. 
    Abstract ( 724 )   PDF (1892KB) ( 309 )   Save
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    Objective   To evaluate the effects of hydrogen sulfide (H2S) on high glucose-induced inflammation of endothelia cells and to explore the possible mechanisms.  Methods   Human umbilical vein endothelial cells(HUVECs)were randomly divided into 4 groups:  group A (5.5mmol/L glucose), group B (16.7mmol/L glucose), group C (16.7mmol/L glucose+100μmol/L NaHS) and  group D (5.5mmol/L glucose+100μmol/L NaHS). After treating for 24h, the expression of nuclear factor-κB (NF-κB) was determined by Western blot,  mRNA expression was examined by RT-PCR, and the activation was detected by electromobility shift assay (EMSA). After treating for 48 h,  protein and mRNA expressions of cyclooxygenase-2 (COX-2) were determined by Western blot and RT-PCR, respectively. Results   ① Compared with that of group A, COX-2 protein expression was distinctly increased(P<0.01)and the mRNA level was also increased in group B(P<0.01). However, COX-2 protein expression was distinctly decreased(P<0.05) as well as mRNA level(P<0.01) in group C when compared with that of group B. ② Compared with that of group A, cytoplasmic NF-κB protein expression was obviously decreased in group B (P<0.05), but NaHS could stabilize the cytoplasmic NF-κB protein level (P<0.01). There were no significant differences between group A to group D in NF-κB mRNA expression. ③ High glucose increased NF-κB activation in group B. Compared with that of group B, decrease in NF-κB activation was observed in group C. There were no differences between group A and group D in NF-κB and COX-2 expressions and NF-κB activity. Conclusion   H2S can protect against high glucose-induced inflammation in human umbilical vein endothelial cells, which may be associated with the suppression of COX-2 expression.

    Effects of erlotinib combined with celecoxib on growth and angiogenesis of human lung cancer xenograft model
    DONG Xue-li, MU Xiao-yan, LIU Qing-liang, SUN Jie
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  49-52. 
    Abstract ( 686 )   PDF (1172KB) ( 345 )   Save
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    Objective   To explore the effects of erlotinib combined with celecoxib on the tumor growth and angiogenesis of lung cancer xenografts in nude mice. Methods   Human lung cancer cell A549 was subcutaneously injected to establish the nude mice xenograft model. The nude mice with tumor were randomly divided into four groups:the control group, the celecoxib group, the erlotinib group and the combined group.  All the mice were given the gavage administration. The status of the mice were observed everyday , and the volume of the tumor was measured twice a week to obtain the tumor growth curve. The mice were put to death to collect the tumor tissues after 40 days. The microvessel density (MVD) was detected by the immunohistochemistry analyses and the expressions of Bcl-2 and Bax were detected by Western Blotting. Blood samples were collected to detect the sVEGF level by ELISA. Results   The tumor volumes of the combined group were significantly smaller compared with the control group, the celecoxib group and the erlotinib group(P<0.05). The microvessel density and the sVEGF level of the combined group were decreased compared with the control group, the celecoxib group and the erlotinib group(P<0.05). The Bcl-2 level of the erlotinib group was lower compared with the control group. There was no significant difference of the Bcl-2 level between the celecoxib group and the control group (P>0.05). The Bcl-2 level of the combined group is lower compared with the control group, the celecoxib group and the erlotinib group(P<0.05). There was no significant difference of the Bax level among the control group, the celecoxib group and the combined group(P>0.05). The value of Bcl-2/Bax of the combined group was lower compared with the celecoxib group and the erlotinib group(P<0.05). Conclusion   Erlotinib combined with celecoxib  have more effective growth inhibition on the human lung cancer xenograft model than erlotinib or celecoxib separately. The possible mechanism may be related to the decrease of  microvessels and the increase of  tumor apoptosis.

    Treatment of atrial fibrillation with radiofrequency modified maze Ⅲ and ganglionic plexus ablation
    CHENG Xian-feng, LIANG Fei, ZHANG Qian, ZHU Xiao-long, ZHANG Hai-zhou, ZHANG Tao, HAN Bo, ZOU Cheng-wei
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  53-56. 
    Abstract ( 914 )   PDF (1147KB) ( 443 )   Save
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    Objective   To investigate the process, effectiveness and influence factors of radiofrequency modified maze Ⅲ(RFMM Ⅲ) and ganglionic plexus(GP) ablation in the patients with atrial fibrillation (AF) undergone open-heart surgery. Methods   Retrospective analysis was used to analyze the clinical data of 102 patients who undergone RFMM Ⅲ and GP ablation treatment of valvular heart disease, congenital heart disease and coronary atherosclerosis sclerotic heart disease with AF from December 2009 to December 2011. The regular follow-up was performed for 6 months after the operation. Results   1 patient died of the low cardiac output syndrome. Re-thoratomy for homeostasis occurred in 3 cases and incision infection occurred in 2 cases. Electrocardiogram showed normal rhythm without AF after the operation (sinus rhythm 93.1% and junctional rhythm 6.9%). After 6-month follow-up, the maintenance rate of sinus rhythm was 84.2%, junctional rhythm occurred in 6 patients (6.3%), and AF recurred in 9 patients (9.5%). Left atrial diameter and history of AF were the risk factors of the recurrence after RFMM Ⅲ and GP ablation(P=0.027 and 0.040 respectively). After radiofrequency ablation in the congenital heart disease and the acquired heart disease, the recurrence rate of AF had no statistical difference (P=0.686). Conclusion   RFMM Ⅲ and GP ablation was more effective and easy to operate, which could improve the successful rate of ablation. This operation had certain innovations and good prospects for the AF treatment, which deserved more clinical applications and long-term follow-up to enhance the long-term efficacy.

    Correlations between levels of plasma  homocysteine and hydrogen sulfide and cerebral circulation dynamics in H type hypertension patients
    ZHAO Qian1, LIU Rui2, XU Zhen-xing1, ZHU Shi-ming1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  57-60. 
    Abstract ( 871 )   PDF (870KB) ( 308 )   Save
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    Objective   To investigate the correlations between plasma homocysteine (Hcy) and hydrogen sulfide (H2S) levels and parameters of cerebral circulation dynamics in H type hypertension patients. Methods   104 patients were graded in H type hypertension group and hypertension group according to the level of plasma Hcy with 32 normal controls as the control group. The levels of plasma Hcy and H2S were measured by specific protein instrument and sulfur sensitive electrode. The parameters of cerebral circulation dynamics were detected by cerebral circulation dynamics detector. The levels of plasma folic acid and vitamin B12 were measured by auto DELFIA analyzer. Results   Compared with those of the controls and the hypertension group, the levels of plasma H2S, folic acid and vitamin B12 significant decreased in the H type hypertension group(P<0.05), and the parameters of cerebral circulation dynamics had significant differences in the experiment group(P<0.001). The level of plasma Hcy in the H type hypertension group had significantly nagative correlations with H2S, folic acid, vitamin B12, Vmean and Qmean (P<0.05).But it had significantly positive correlations with Rv, Zc and DR (P<0.05). The level of plasma H2S in the experimental group had significantly positive correlations with Qmean (P<0.05) and nagative correlations with Rv (P<0.05). Multiple linear regression analysis revealed a significant and independent association of Qmean with the increasing level of plasma Hcy, as was the association of Rv with the decreasing level of plasma H2S. The increasing level of plasma Hcy was an independent risk factor for the decreasing cerebral blood flow, and the decreasing level of plasma H2S was an independent risk factor for the increasing cerebral vascular resistance. Conclusion   Brain vascular physiology function was affected by the levels of plasma Hcy and H2S, which can cause changes of brain hemodynamics in H type hypertension.

    Relationship between matrix metalloproteinases-9 gene polymorphism and the cardiovascular remodeling in  hypertension patients
    XU Zhen-xing1, LIU Rui2, ZHAO Qian1, ZHANG Jun-sheng3, ZHU Shi-ming1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  61-64. 
    Abstract ( 768 )   PDF (865KB) ( 290 )   Save
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    Objective   To explore  the relationship between matrix metalloproteinases-9 gene polymorphism and the cardiovascular remodeling in  hypertension patients. Methods   53 hypertension patients were randomly selected. 30 healthy people were selected as the control group. Ventricular end-diastolic interventricular septum (IVSTd), end-diastolic left ventricular wall thickness(LVPWTd) and carotid intima media thikness(IMT) were determined by Doppler ultrasonography. The blood pressure of upper and lower limbs was measured and the ABI was calculated. The C-1562T gene polymorphism of MMP-9 was measured by polymerase chain  reaction combined with restriction enzyme digestion. Results   MMP-9 gene polymorphism was not associated with the onset and development of hypertension(P>0.05);IVSTd, LVPWTd and IMT of hypertension patients who carried the T alleles was thicker than that of the hypertension patients who didn’t carry the T alleles(P<0.05);ABI of hypertension patieints who carried the T alleles was lower than that of the patients who didn’t carry the T alleles;The C-1562T gene polymorphism of the hypertension patients was positively correlated with the IVSTd, LVPWTd and IMT(P<0.05),and was negatively correlated with the ABI(P<0.05). Age had negative correlations with the ABI in the  hypertension patients. Conclusion   C-1562T gene polymorphism of MMP-9 gene is one of the important factors that effect the cardiovascular remodeling in the hypertension patients.

    Deletion of the RASSF1A gene and promoter methylation in the bone marrow of patients with myelodysplastic syndromes
    XU Jia, SONG Qiang
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  65-69. 
    Abstract ( 816 )   PDF (1097KB) ( 251 )   Save
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    Objective   To examine the expression level  of   RASSF1A gene and the methylation status of its promoter region in  bone marrow of patients with myelodysplastic syndromes (MDS), and to explore the correlation between RASSF1A gene methylation pathogenesis and development of MDS, in order to provide a new target for demethylation drugs. Methods   30 patients with MDS were chosen as the study group and 15 patients with non-malignant hematological diseases were chosen as the control group. Divide the study group according to the MDS IPSS risk score.Methylation specific PCR(MS PCR) was employed to detect the methylation status of the RASSF1A gene promoter region in the 30 patients with MDS. Expression levels of RASSF1A mRNA were also determined by reverse transcription PCR (RT PCR).Contrast the RASSF1A methylation status of the MDS patients before and after using Decitabine. Results   The RASSF1A methylation rate of the study group (73.33%) was much higher than that of the control group(P<0.05), and the RASSF1A methylation rate of the medium-high risk MDS patients(81.8%) is obviously higher than that of the low-risk MDS patients(18.2%)(P<0.05);Meanwhile,the RASSF1A mRNA expression level of the study group was significantly lower than that of the control group(P<0.05).The RASSF1A methylation level of the MDS patient after Decitabine treatment was lower than that of newly diagnosed patients. Conclusion   A significant correlation existed between RASSF1A hypermethylation and the loss of expression of RASSF1A mRNA in MDS. The hypermethylation of the RASSF1A gene promoter region may be one of the mechanisms related to the pathogenesis of MDS. Decitabine can reduce the methylation levels in the RASSF1A gene and it also provides a new theoretical basis for using Decitabine as the treatment of MDS.

    Clinicopathological significances of WIF-1 gene methylation and Wnt-5a protein expression in chondrosarcoma
    HOU Bei-bei, WANG Yan, ZHANG Juan, LIANG Xiao-hui, LI Jin-song
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  70-74. 
    Abstract ( 999 )   PDF (1580KB) ( 255 )   Save
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    Objective   To study the promoter methylation status of Wnt inhibitory factor-1 gene (WIF-1) and the protein expression of Wnt-5a in chondrosarcoma and then explore their clinicopathological significances. Methods   Methylation specific PCR(MSP) and immunohistochemical SP method were used respectively to investigate the methylation of WIF-1 gene and the expressions of Wif-1 protein and Wnt-5a protein in 61 chondrosarcoma samples and 40 matched chondroma samples. Results   The methylation of WIF-1 gene and the expression of Wnt-5a protein obviously increased in chondrosarcoma compared with chondroma(P<0.05). The expression of Wnt-5a protein was significantly higher in tumors with infiltration and/or metastasis than in that without any invasion and metastasis (P<0.05). The methylation of WIF-1 gene was related to the decreased expression of Wif-1 protein and the over-expression of Wnt-5a protein (KappaWif-1 Value 0.747,PWif-1<0.001;KappaWnt-5a Value 0.468, PWnt-5a<0.05). Conclusion   WIF-1 gene methylation may play an important role in the early stage of chondrosarcoma formation. The over-expression of Wnt-5a protein is positively related to the invasiveness of chondrosarcoma. Synergistic effects might exist between WIF-1 gene methylation and over-expression of Wnt-5a protein in the oncogenesis of chondrosarcoma.

    Application of intracranial pressure monitoring in patients with severe traumatic brain injuries
    LI Zhi-qiang1, DAI Xin-lian1, SHEN Dong-qing1, ZHAO Jun1, LIU Hong-wei1,2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  75-78. 
    Abstract ( 856 )   PDF (861KB) ( 363 )   Save
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    Objective   To explore the application value of intracranial  pressure(ICP)monitoring in patients with sever traumatic brain injuries (TBI). Methods   The relative data of 142 ICP monitoring patients  were compared to those of 146 non-ICP monitoring patients, especially on important complications and Glasgow outcome scale (GOS). Results   ICP was slightly,moderately and severely elevated in 43,69 and 30 cases in the monitoring group respectively. They received corresponding proper treatment. Complications such as electrolyte imbalance and renal damage in non-monitoring group were more than in monitoring group. GOS score was significantly higher in monitoring group than in non-monitoring group. Conclusion    The continuous ICP monitoring can demonstrate intracranial pressure change and help to guide the correct clinical nursing and treating, and so as to improve the clinical outcome.

    Clinical study of the related  risk factors associated with endometriosis 
    YU Ling, TIAN Yong-jie
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  79-83. 
    Abstract ( 785 )   PDF (875KB) ( 312 )   Save
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    Objective   To investigate the  hospitalbase incidence of endometriosis among women with different ages and the risk factors associated with endometriosis.  Methods   A case control study was carried out in Jinan including 346 endometriosis patients and 346 healthy people. The data were collected and analyzed with single factor analysis and multivariate unconditional logistic regression analysis among age, individual aspects, menstrual characteristics and reproductive activity. Results   ① The age peak of incidence of endometriosis was between 25 and 50. ② Single factor analysis showed that occupation as cadre and mechanics, college educated, A and O blood types, menarche before or just at age 13, menstrual cycle <27days,menstruallengths >or=8 days, dysmenorrhea, one live birth, abortion numbers >or=2, infertility and intrauterine device use were identified as risk factors. There were significant differences between the two groups(P<0.05). ③ Multifactor analysis confirmed that the risk factors associated with endometriosis included early menarche, short cycles, long menstrual lengths and more abortions numbers. Pregnancy number and labor number were proved to be the protective factors. Conclusion   Endometriosis seemed to be related to early menarche, short cycles, long menstrual lengths, and more abortions numbers.

    Dehydroepiandrosterone supplementation improve in vitro fertilization outcome among women with poor response
    LU Qun, SHEN Huan, WANG Cong, ZHENG Xing-bang, CHEN Xi, LIANG Rong, WEI Li-hui
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  84. 
    Abstract ( 799 )   PDF (905KB) ( 402 )   Save
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    Objective   To evaluate the effect of treatment with dehydroepiandrosterone ( DHEA ) on in vitro fertilization and embryo transfer (IVF-ET) outcomes among women with poor response. Methods   Thirty-seven poor response women with at least one IVF cycles received 5075mg of DHEA daily for one to three months. Antral follicle count (AFC), basal follicle stimulating hormone (FSH),luteinizing hormone (LH), estradiol (E2), testosterone (T), free T (FT), DHEA and IVF outcome parameters were compared before and after the DHEA treatment by paired t-test. Results   Women received (52.03±12.33)mg of DHEA daily for an average of (84.05±35.31) days. Paired analysis of endocrine parameters in 37 patients showed that AFC, T, FT and DHEA increased after DHEA supplementation (P<0.01), while FSH and LH levels decreased. IVF cycle outcomes demonstrated significant increases on retrieved oocytes and high quality embryos after the DHEA treatment. Ten women conceived (clinical pregnancy rate, 27.03%) and eleven healthy babies were born. Conclusion   DHEA supplementation could significantly increase the androgen level in women with poor response, which could improve ovarian reserve, and increase the number of retrieved oocytes and pregnancy rate.

    Treatment of Vancouver type B3 periprosthetic femoral fracture with greater trochanter reattachment device and cortical strut bone allograft
    WANG Chao, ZHANG Wei, GUAN Da-wei, LI Wei, WANG Xian-quan, WANG Wen-hao
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  88-92. 
    Abstract ( 771 )   PDF (1363KB) ( 382 )   Save
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    Objective   To analyze the therapeutic effect of  Vancouver type B3 periprosthetic femoral fracture after treating with greater trochanter reattachment device (GTR) and cortical strut bone graft in the prosthesis revision of the total hip arthroplasty.  Methods   A total of 26 patients were selected from January 2006 to January 2011, including 5 males and 21 females, aged 55~79 years, 63.52 years on average. All the cases were treated by the Zimmer greater trochanter reattachment device (GTR), and the Zimmer Wagner uncemented long stem. Harris score was used to evaluate the hip functions before and after the operation.  Prosthesis stability and the bony union were assessed by the digital radiography.  Results   All the patients were followed-up for 25 months on average (12~35 months) after the operation. All the fractures reached union, and there was no infection, stem loosening, nonunion and malunion. The Harris score was (35.26±3.35) before the operation, which was improved to (84.70±6.21) after the operation, and the good-to-excellent rate was 80.77% (5 excellent cases, 16 good  cases and 5 average cases). X-ray showed that the prosthesis was stable, and the host bone and bone graft achieved bony union. Conclusion   The prosthesis revision of the total hip arthroplasty with the greater trochanter reattachment device (GTR) and cortical strut bone graft used for the Vancouver type B3 periprosthetic proximal femur fractures has the advantages of simple manipulation, less complications, good recovery of the hip function and satisfactory results in the short period.

    Association of TLR7/8 polymorphisms with risk and severity of asthma in Han population of Jiangsu province of China
    ZHANG Qian1,2, QIAN Fen-hong3, ZHOU Lin-fu4, WEI Guo-zhen1,BAI Jian-ling5, YIN Kai-sheng4, SHI Yi2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  93-98. 
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    Objective   To evaluate the effects of polymorphisms in TLR7 and  TLR8 on asthma risk and asthma-related phenotypes in Han population of Jiangsu province of China. Methods   We consecutively recruited 318 unrelated adult asthmatic patients and 352 healthy volunteers of Han population from the same area in Jiangsu province. Genotyping of the selected SNPs, TLR7 rs5935436, TLR8 rs3761623 and rs3764880 was performed using multiplex PCR combined with tagged array single base extension technology(SNPstream). Total IgG level in serum was determined by ELISA. We conducted case-control and case-only association studies between the selected SNPs in TLR7 and TLR8 and asthma or asthma-related phenotypes. Results   The T allele of rs5935436 SNP in TLR7 was a protective factor for males in developing asthma(adjusted OR=0.126, 95%CI=0.016-0.995, P=0.049). The G allele of the rs3761623 appeared more frequently in male asthmatics without allergic rhinitis (adjusted OR=3.782, 95% CI=1.02214.045, P=0.047), while the CT/TT genotype of the rs5935436 appeared less frequently in female asthmatics with allergic rhinitis (adjusted OR=0.180, 95%CI=0.041-0.902, P=0.037). The homozygote AA of rs3761623 and GG of rs3764880 were positively associated with lower FEV1% and asthma severity in female asthmatics. These results were confirmed by haplotype analysis. Conclusion   TLR7/8 polymorphisms may play an important role in the pathogenesis of asthma, which is gender-dependent. This could be clinically useful to identify patients at risk of asthma and prevent the occurrence of asthma.

    Correlation of Trefoil factor 3 expression in gastric carcinoma and precancerous lesions with angiogenesis
    WANG Chen-chen1, GU Xiao-meng1, WANG Yan2, HUANG Shan-ying3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  99-103. 
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    Objective   To investigate the expression of TFF3 in gastric carcinoma and precancerous lesions, and explore the correlation in gastric carcinogenesis and angiogenesis. Methods   The expression of TFF3 was detected by  immunohistochemistry in 26 cases of superficial gastritis, 25 cases of intestinal metaplasia, 20 cases of atypical hyperplasia and 66 cases of gastric carcinoma. MVD was detected by immunohistochemistry using anti-CD34 antibody. Results   The expression of TFF3 was lower in gastric carcinoma than in intestinal metaplasia and atypical hyperplasia, with higher expressions than that in superficial gastritis(P<0.01); MVD was higher in gastric carcinoma than other three groups(P<0.01). There was no significant difference of MVD between intestinal metaplasia and atypical hyperplasia(P>0.05), with higher expressions than that in superficial gastritis(P<0.01). The expression of TFF3 was correlated with lymph node metastasis and TNM staging(P<0.05). The expressions of TFF3 and MVD were positively correlated (r=0.672). MVD with positive expression of TFF3 was higher than that with negative expression(P<0.01). Conclusion   TFF3 may play important roles in carcinogenesis , invasion, metastasis and angiogenesis. It can be used as a useful indicator of the early prediction and prognosis of gastric cancinoma.

    Type D personality in patients with hemiplegic stroke in the rehabilitation phase
    CHEN Jin1, QI Zhen-hua1, JIANG Hong1, ZHANG Can-can1, JIANG Ping-yin2, DANG Hong-mei2, ZHAO Peng2, ZHANG Hong-jing1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  104. 
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    Objective   To investigate the distribution of type A  personality  and type D personality in patients with hemiplegic stroke in the rehabilitation phase, and the influence factors of the two kinds of personalities in stroke. Methods   77 patients were investigated with the Type A  Scale-Chinese(TAS-C) , Type D Scale-14(DS14), Self-Rating Depression Scale(SDS), Self-Rating Anxiety Scale(SAS)  and Life Orientation Test-Revised(LOT-R) on the second day of admission to judge the personality and mood states of the patients. The correlation was analyzed. 58 healthy individuals were used as the control. Results   The detection rates of type A personality were 70.1% in the study group and 24.1% in the control group, while the detection rates of type D personality were 67.5% in the study group and 25.9% in the control group,and the D&A were 44.2% in the study group and 8.6% in the control group (all P<0.01). The scores of the study group were significantly higher compared with the control group in Type A score, TH, CH and Type D score,  NA, SI (all P<0.01).The regression analysis showed that type A personality, type D personality and NA,SI were risk factors of the stroke( all P<0.01). TH,CH and NA were positively correlated with SAS total scores(r=0.25-0.34;all P<0.05). NA and SI were negatively correlated with the scores LOT-R( all P<0.05), and positively correlated with SDS total scores(all P<0.05). The score of NA had significant correlation with depression and anxiety(P<0.01). The scores of TH were correlated with the optimism(P<0.05). The scores of SI were correlated with SAS total scores(P<0.01). Conclusion   Type D personality is closed to the hemiplegic stroke, and could incur an increased risk of anxiety and depression.

    Correlation between the psychological health status and the coping style in the female prisoners
    SHI Liang1, TANG Mao-qin2, HU Lei2, HE Xu3, ZHAO Hong-yan4
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(2):  108-112. 
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    Objective   To investigate the female prisoners’ psychological health status, the coping style and the relevant affecting factors involved, providing a scientific basis for further psychological education. Methods   The 1500 female prisoners of Shandong Female Prison were investigated with the Coping Style Questionnaire (CSQ), Symptom Checklist  90(SCL-90)  and the General Status Questionnaire, with the relevant affecting factors analyzed.  Results   It was found that 1420 female criminals were analyzed, 45.7% female prisoners had the psychological health problems, with 36.1%  being medium or severe. Scores of all the factors were higher in prisoners than those in the normal controls. The differences were statistically significant(t1-t11=2.623-7.015, P1-P11<0.01). The psychological health status was positively correlative with factors like problem solving,illusion and help seeking(r=0.019-0.183, P<0.05), and it was negatively correlative with factors like self-reproach, retreat and rationalization(P<0.05).  The primary factors of influence female criminals’s psychological health status were the prison term and positively solution style (t=-2.729-4.510,P<0.05).  Conclusion   The results indicate that the female criminals have obviously psychological problems. The psychological health status of female prisoners is  correlative closed with the coping style.