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Table of Content

      
    10 March 2013
    Volume 51 Issue 3
    Articles
    Effects of NAD on the mRNA expression of collagen  type I induced by Angiotensin II in cardiac fibroblasts
    ZHAO Li-xiang, WU Xin-ning, WANG Xi, LI Na, CHEN Tong-shuai, LIU Jun-ni, BU Pei-li
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  1-5. 
    Abstract ( 1063 )   PDF (2585KB) ( 1242 )   Save
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    Objective   To explore the effect of NAD on the mRNA expression of collagen type I induced by angiotensin II in cardiac fibroblasts in vitro.Methods   Neonatal rat cardiac fibroblasts  were isolated from wistar rats and then cultured. The generation 2-4 were used for the experiment and treated as follows: control group, Ang II (0.01, 0.1,  1μmol/L) group,NAD (250μmol/L )group, Ang II (1μmol/L)+ NAD (250μmol/L )group, FAM group, Sirt1-siRNA+AngII (1μmol/L) group, Sirt1-siRNA group, Sirt1-siRNA+NAD(250μmol/L)group and Sirt1-siRNA+ Ang II (1μmol/L)+NAD(250μmol/L)group.  24 hours later, the mRNA expressions of SIRT1 and collagen I were measured by real time reverse transcription-polymerase chain reaction(Real time RT-PCR). Results   After treatment with Ang II for 24 hours,the mRNA expression of collagen I was enhanced in a Ang II dose-dependent manner(P<0.05). In contrast with control group, the mRNA expression of SIRT1 was enhanced after being treated with NAD(250μmol/L)(P<0.05) and the mRNA expression of collagen I in AngII (1μmol/L)+ NAD (250μmol/L) group was attenuated compared with Ang II (1μmol/L) group (P<0.01).  Compared with FAM group, SIRT1 mRNA expression was attenuated(P<0.05) and collagen I mRNA expression was increased (P<0.05) in the other groups.Conclusion   The results suggest that NAD and SIRT1 may be beneficial to cardiac fibrosis by attenuating the mRNA expression of collagen type I induced by Ang II in cardiac fibroblatsts.

    Effects of troglitazone on the expressions of CD147,  TGF-β1 and MMP-9 in GH3 cells
    LIU Bo, YANG Wei, MU Cheng-zhi, ZHANG Yu-bao, XU Guang-ming
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  6-9. 
    Abstract ( 991 )   PDF (1208KB) ( 528 )   Save
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    Objective   To explore the effect of troglitazone(TGZ), a ligand for peroxisome proliferator-activated receptor γ(PPAR-γ), on the expressions of cluster of differentiation 147(CD147), transforming growth factor beta 1(TGF-β1) and matrix metalloproteinase 9(MMP-9) in rat GH-secreting GH3 cells. Methods   GH3 cells were devided into control group and experimental groups (1×10-3, 1×10-4 and 1×10-5 mol/L TGZ). After treated with TG2 for 48 hours, expressions of CD147, TGF-β1 and MMP-9 at protein and mRNA levels were detected by Western blot and Real-time PCR respectively. Results   The expressions of CD147, TGF-β1 and MMP-9 at protein and mRNA levels in all the TGZ treated groups showed significant differecnces compared with control group(all P<0.05). Moreover, expressions of the above three indicators decreased both at protein and mRNA levels when TGZ concentration was increased. Conclusion   TGZ can inhibit expressions of CD147, TGF-β1 and MMP-9, which   might relieve the invasiveness of GH3 cells.

    Expression of TIPE2 and its association with Aβ1-40 deposition in brain tissue of type 2 diabetic GK rat
    SUN Zhong-wen1, DU Juan2, BAO Meng-xin3, CHEN Juan1, ZHU Mei-jia4
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  10-14. 
    Abstract ( 1303 )   PDF (3722KB) ( 552 )   Save
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    Objective   To investigate the expression of tumor necrosis factor-α induced protein-8 like-2 (TIPE2) and its association with amyloid beta 1-40 deposition in brain tissue of type 2 diabetic GK rat. Methods    GK rats were selected as type 2 diabetic rat model (GK group), and Wistar rats were selected as non-diabetic rat model (Wistar group). Then the spatial distribution of TIPE2 and Aβ1-40 in brain tissue was observed by immunohistochemistry (IHC). The mRNA and protein level of TIPE2 in brain tissue was detected by RT-PCR and Western blot respectively. The expression of Aβ1-40 in brain tissue was quantized by ELISA. Results   TIPE2 was observed in neuroglial cells and small blood vessels of cerebral cortex and subcortical structures in rat brain tissue. Compared to Wistar group, the expression of TIPE2 in brain tissue was significantly higher in GK group (P<0.05). Aβ1-40 was detected in neurons, neuroglial cells, and brain blood vessel wall of cerebral cortex and hippocampus. Compared to Wistar group, expression of Aβ1-40 in brain tissue was significantly higher in GK group (P<0.05). The expression of TIPE2 was positively associated with Aβ1-40 level both in Wistar group(r=0.951, P<0.05) and GK group (r=0.954, P<0.05). Conclusion   TIPE2 expression in brain from type 2 diabetic GK group increases significantly than Wistar control group and is positively associated with Aβ1-40 deposition, indicating the role of TIPE2 in diabetic brain damage.

    Expression of HBV and its effects on the transdifferentiation in HK-2 cells
    XU Hui1, WANG Yi-guo2, LIU Chang-hong2, LIU Qian2, DU Juan3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  15-20. 
    Abstract ( 1110 )   PDF (2566KB) ( 507 )   Save
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    Objective   To investigate the expression of HBV and effects on the transdifferentiation in HK-2 cells. Methods   HK-2 cells were cultured in vitro and we devided them into the HK-2 group,  the HK-2-PHY106-HBV group (HK-2 cells transfected with PHY106-HBV plasmid) and the HK-2-PHY106 group (HK-2 cells transfected with PHY106 control plasmid). HK-2 cells were transfected by lipofectamineTM2000. Supernatant of culture was collected for examining HBsAg and HBeAg by using ELISA; The expression of E-cadherin and α-smooth muscle actin in HK-2 were assayed by immunocytochemistry and Western blot after the cells were transfected for 72h. TGF-β1 mRNA was detected by RT-PCR after the cells were transfected for 72h. Results   The HBsAg and HBeAg were detected with a higher expression in the HK-2-PHY106-HBV group. The E-cadherin detected by immunocytochemistry and Western blot showed a significantly lower expression in the HK-2-PHY106-HBV group(P<0.05). However, the expression of α-SMA in the HK-2-PHY106-HBV group was higher than that of the other two groups(P<0.05). Additionally,  we also found TGF-β1 mRNA expression was upregulated in the HK-2-PHY106-HBV group showed by RT-PCR(P<0.05).  Conclusion   HBV can be replicated highly efficiently in HK-2 cells and can make HK-2 cells occur transdifferentiation. The mechanism may be by upregulating TGF-β1 expression.

    Mechanisms of EGCG inhibiting the proliferation of human colon cancer HT-29 cells and preventing angiogenesis
    XIN Yu1, YUAN Jun-hua2, SUN Cheng-gang1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  21-26. 
    Abstract ( 1067 )   PDF (1309KB) ( 1208 )   Save
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    Objective   To explore the mechanisms of epigallocatechin-3-gallate(EGCG) inhibiting the proliferation of human colon cancer HT-29 cells and preventing the angiogenesis in vitro. Methods   In vitro,  human colon cancer cell line HT-29 was treated by EGCG in various concentrations(0, 25,  50,  100μg /mL) for 24h. The inhibitory effect of proliferation was assayed by MTT method. The mRNA and protein expressions of transforms growth factorsβ1(TGF-β1),  15-hydroxyprostaglandin dehydrogenase (15-PGDH),  cyclooxygenase-2(COX-2)and vascular endothelial growth factor(VEGF) in human colon cancer cell line HT-29 were respectively examined by RT-PCR and Western blot. Results   MTT assay showed that EGCG inhibited HT-29 cells in a dose-and-time dependent manner(P<0.05). It was found that the expressions of TGF-β1 and 15-PGDH mRNA and protein  were up-regulated and COX-2 and VEGF were down-regulated,  both in a dose-dependent manner(P<0. 05). Conclusion   EGCG inhibits the proliferation of HT-29 cells in a dose-and-time dependent manner. EGCG possibly prevents angiogenesis of HT-29 cells via activating TGF-β1/15-PGDH pathways and inhibiting COX-2 and VEGF pathways in a dose-dependent manner.

    Effect of losartan on the rat model of experimental hepatic fibrosis
    YANG Yan1, YAN Chun-ying1, CHEN Xu2, LIN Xiao-yan3, WANG Lai-cheng4, SHI Jun1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  27-31. 
    Abstract ( 997 )   PDF (3474KB) ( 344 )   Save
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    Objective   To explore the effect of angiotensin Ⅱ receptor blocker losartan on rats hepatic fibrosis induced by carbon tetrachloride (CCL4). Methods   Hepatic fibrosis was induced in rats by intraperitoneal injection of carbon tetrachloride(40%). Meanwhile, losartan was given to the fibrosis+losartan intervention group rats(daily gavage), and sterile saline was given to the control rats for 6 weeks. Blood and liver tissue samples were taken for detecting aspartate aminotransferase (AST) and alanine aminotransferase (ALT).The pathological changes and fibrosis extent of liver tissues were observed by light microscope and polarizing microscope respectively. Collagen type I and collagen type Ⅲ were detected by immunohistochemistry.Liver AngⅡ and AT1R mRNA expressions were determined by RT-PCR. Results   Compared with those of the model group, liver inflammation and hepatic fibrosis were significantly reduced in the fibrosis+losartan intervention group;collagen type I and collagen type Ⅲ content were lowered in the fibrosis+losartan intervention group(P<0.05); the liver function was improved significantly.Meanwhile,the levels of AngⅡand AT1R mRNA  in liver tissues also decreased(P<0.05). Conclusion   Losartan may has a therapeutic effect on carbon tetrachloride-induced liver fibrosis rats by reducing the expressions of liver AngⅡand AT1R.

    Expression of  GDNF in LPS-treated mouse Sertoli cells and its effects on spermatogonial stem cells
    SHI Kun, ZHANG Xiao-li, ZHANG Hai-yu, JI Shu-fang, HAO Jing
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  32-41. 
    Abstract ( 1187 )   PDF (7408KB) ( 449 )   Save
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    Objective   To investigate the changes of GDNF expression in sertoli cells(SCs) of LPS-treated Kunming(KM) mice and its effects on spermatogonial stem cells (SSCs). Methods   SCs were isolated and purified from testes of 5~6 weeks old KM mice. The SCs were cultured and divided into four groups: the normal culture medium(group con), the medium with LPS treatment 1 day( group 1d ), treatment 2 days( group 2d) and treatment 3 days( group 3d). The medium of four groups were collected respectively as the conditioned medium. RT-PCR and Western blotting were performed to investigate the changes of GDNF expressions of four groups SCs. The SSCs from the testes of 5~7days KM mice were isolated and cultured, and divided into four groups: con, groupⅠ, group Ⅱ and group Ⅲ. The SSCs were cultured for 96 hours in the conditioned medium respectively. Immunofluorescent staining was performed on SSCs to investigate the expressions of PLZF, oct4, PCNA, c-kit and sohlh2. RT-PCR was performed to investigate the expressions of Gfrα1, c-ret, Bcl6b and Etv5. Results   RT-PCR and Western blotting results revealed that the expression of GDNF in SCs was decreased in the order of group con, group 1d, group 2d and group 3d. Immunofluorescence and RT-PCR results revealed that the expressions of PLZF, oct4 and PCNA in SSCs were decreased in the order of group con, group 1d, group 2d and group 3d, and the expressions of c-kit and sohlh2 were increased in the order of group con, group 1d, group 2d and group 3d. RT-PCR results revealed that the expressions of Gfrα1, c-ret, Bcl6b and Etv5 in group 1d, group 2d and group 3d were decreased compared with group con. Conclusion     The expression of GDNF in LPS-treated mouse SCs is decreased. The differentiation and self-renewal of SSCs can be stimulated and inhibited respectively by the reduction of GDNF expression in SCs via down-regulation of Bcl6b and Etv5 expression.

    Effects of N-acetylcysteine  on resistance to paclitaxel in cervical carcinoma cells
    SUN Gao-ying1, WANG Xiao-yuan2, ZHANG Xiao-jin1, HAO Xiu-yu1, GUO Yuan-fang1, WANG Xiao1, BI Wen-xiang1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  42-47. 
    Abstract ( 1161 )   PDF (1238KB) ( 561 )   Save
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    Objective   To construct a paclitaxel (PTX)-resistant human cervical carcinoma cell line and observe the effects of N-acetylcysteine (NAC) on resistance to PTX in the cells.  Methods   Hela cells were treated with PTX to construct the PTX-resistant Hela/PTX cell line. The cell shape, cell growth curves and drug resistance index as well as the cellular redox state and the taxol resistance gene 1 (Txr1) expression were observed or determined. Hela/PTX cells were treated with NAC, and the effects of NAC on resistance to PTX in the cells were observed.  Results   After Hela cells were treated with PTX for 10 months, the PTX-resistant cervical carcinoma Hela/PTX cells could be obtained, which grew well in 500μg/L of PTX. Compared with Hela cells, the size of Hela/PTX cells was larger, and there were more cytoplasmic granules in them. The population doubling time for Hela/PTX cells was 1.32 times of that for Hela cells. Hela/PTX cells manifested a stronger resistance to PTX than Hela cells did with the resistance index of 122.69. They had higher levels of reactive oxygen species (ROS) and Txr1 mRNA, a lower level of reduced glutathione(GSH), and lower activities of superoxide dismutase, catalase and glutathione peroxidase than Hela cells. There was no significant difference between Hela/PTX and Hela cells in the level of oxidized glutathione (GSSG) or the GSH to GSSG ratio. After Hela/PTX cells were treated with NAC, the levels of ROS and Txr1 mRNA in them decreased, and they were more sensitive to PTX.  Conclusion   In Hela/PTX cells, which resisted to PTX, there was a redox imbalance, and the Txr1 transcripts increased. NAC could reduce the levels of ROS and Txrl mRNA in Hela/PTX cells and reversed the sensitivity of them to PTX.

    Effects of human umbilical cord mesenchymal stem cells on nerve fibers in the lesions of rat endometriosis  model
    CHEN Yan1, LI Dong2, ZHANG Zhe1, SHI Qing2, WANG Da-kun2, LIU Huan2, WANG Guo-yun1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  48-51. 
    Abstract ( 1082 )   PDF (2154KB) ( 384 )   Save
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    Objective   To investigate the influence of human umbilical cord mesenchymal stem cells (hUC-MSCs) on nerve fibers in the lesions of rat models of endometriosis (EMs). Methods   hUC-MSCs were isolated and cultured from fresh human umbilical cord, then identified by flow cytometry. Adult female Wistar rats, weighing 180-210g, were surgically induced EMs. Thereafter the EMs rat models were randomly divided into two groups (12 rats each), namely the treatment group and the control group. Two weeks later, the treatment group received hUCMSCs by injection of normal saline into the tail vein once a week for two weeks. Meanwhile, the control group only received the same volume of normal saline. EMs lesions were collected four weeks later. The specimens were sectioned and stained immunohistochemically with antibodies against Neurofilament (NF) and Growth Associated Protein 43 (GAP-43). Results   Compared to that of the control group, the mean density of NF-immunoreactive nerve fibres of the treatment group was  remarkably lower(P<0.01). Besides, the mean density of nerve fibres stained with GAP-43 was also much lower in the treatment group in comparison with that of the control group(P< 0.01).  Conclusion   MSCs from human umbilical cord could reduce the nervefiber density of EMs lesions.  

    Effects of potassium iodate and bisphenol A on the proliferation of  thyroid carcinoma cells
    LI Ya-nan1, JIANG Ling1, HU Xiu-hui1, ZHANG Xiao-li1, LI jie2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  52-57. 
    Abstract ( 1209 )   PDF (2994KB) ( 560 )   Save
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    Objective   To observe the effects of potassium iodate and bisphenol A(BPA)on proliferation of thyroid carcinoma cells BHP10-3. Methods   10-3~10-10mol/L potassium iodate,  10-3~10-10mol/L bisphenol A and 10-7mol/L potassium iodate +10-8mol/L bisphenol A were added to the culture of BHP10-3 cells. Cell proliferation activity was examined by CCK-8 colorimetric assays,  and the cell cycles of the stimulated cells were analyzed by flow cytometer. The expression of Cyclin D1 protein was observed by western-blot,  and the expression of cyclinD1 mRNA was observed by RT-PCR. Results   Compared with the control group,  the absorbance of the potassium iodate group,  the BPA group and the potassium iodate+BPA group was higher (P<0.05), the proportion of cells in the G0-G1 phase decreased and proportion of cells in the S phase increased significantly (P<0.05), the expression levels of Cyclin D1 protein and mRNA were up-regulated significantly(P<0.05). Conclusion   The potassium iodate and BPA can promote the proliferation of thyroid papillary carcinoma cells in a certain concentration range.

    Inflammatory responses of PGRN-deficient peritoneal macrophage to bacterial lipopolysaccharide in vitro
    LIU Lu1, ZHANG Wen1, CHEN Han-xiang1, ZHENG Lin1, LU Yi2,WANG Hong1, TANG Wei1, ZHAO Wei-ming1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  58-62. 
    Abstract ( 1213 )   PDF (1963KB) ( 548 )   Save
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    Objective   To investigate the effects of progranulin (PGRN) in the inflammatory responses of peritoneal macrophages (PMs) to bacterial lipopolysaccharide (LPS) in vitro. Methods   Peritoneal exudate cells (PECs) were induced and extracted from wild-type (WT) mice and PGRN gene knock-out mice (KO), then the number, morphology and classes of PECs were subsequently evaluated. Surface markers CD11b and F4/80 of PMs were tested by flow cytometry. PMs derived from WT or KO mice were treated with LPS and WT PMs were treated with PBS, LPS, recombinant PGRN or LPS plus recombinant PGRN respectively. Supernatants of cultivation were collected after 24-hours incubation and concentrations of TNF-α, IL-1β, IL-12 and production of NO were detected by ELISA or Griess assay respectively. Results   There were no significant differences in cell number, classes and expression of surface makers CD11b and F4/80 between WT and KO mice-derived PECs. Higher concentration of TNF-α, IL-1β, IL-12 and more NO production were detected in the supernatants of KO PMs stimulated by LPS compared to those of WT PMs. Additionally,recombinant PGRN dramatically inhibited the production of pro-inflammatory cytokines TNF-α, IL-1β, IL-12 and inflammatory intermediate NO of WT PMs stimulated by LPS. Conclusion   PGRN KO PMs display a stronger inflammatory response than WT PMs when treated with LPS. In addition, recombinant PGRN powerfully inhibits LPS stimulating production of TNF-α, IL-1β, IL-12 and NO of PMs.

    Effects of prostaglandin E2-EP4 receptor antagonist on rat experimental autoimmune neuritis
    LU Xiao-li, TAN Xiao-dong
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  63-67. 
    Abstract ( 1131 )   PDF (2490KB) ( 399 )   Save
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    Objective   To explore the role of L161982, one  prostaglandin E2-EP4 receptor antagonist, and its mechanism in rat experimental autoimmune neuritis (EAN). Methods   The rats were injected with bovine peripheral nerve myelin(BPM) antigen emulsion to prepare EAN model. Twenty-one Lewis rats were randomly divided into treatment group A, treatment group B and control group. Treatment group A were intraperitoneally injected daily with L161982(5mg/kg) from one day before immunization to the eighth day after immunization (immunization phase). Treatment group B were intraperitoneally injected daily with L161982(5mg/kg) from the fifth to sixteenth day after immunization (onset phase), and control group were intraperitoneally injected with the same volume of L161982 dissolvent. The effects were assessed in terms of appearance of clinical signs, clinical score, histopathology, and IFN-γ and IL-17 expressions in sciatic nerve sections, and the draining lymph nodes lymphocyte proliferative response was detected by cell counting kit-8(CCK-8). Results   Treatment group A and treatment group B displayed a significant delay in the onset of EAN (P<0.05), a decreased inflammatory cell infiltration into the sciatic nerve(P<0.05), decreased numbers of IFN-γ and IL-17 expressions in the sciatic nerve(P<0.05), and reduced BPMstimulated T lymphocyte proliferative responses(P<0.05) compared to the control group. Only treatment group A had decreased peak clinical score(P<0.05). Conclusion   L161982 ameliorates the severity of EAN, presumably by moderating the over-expression of proinflammatory cytokines and suppressing self-reactive T lymphocyte proliferation. The immunization phase has a more significant therapeutic effect.

    Analysis of linear and nonlinear heart rate variability in patients with essential hypertension and ventricular premature beats
    QI Jia1, MA Yu-ru2, YANG Yong-li3, LI Li1, MA Xiao4
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  68-71. 
    Abstract ( 1336 )   PDF (814KB) ( 430 )   Save
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    Objective   To investigate the changes of linear and nonlinear heart rate variability(HRV)parameters and the correlation between HRV parameters and the number of ventricular premature beats in patients with essential hypertension and ventricular premature beats. Methods   24-hour long-term HRV parameters and 5-minute short-term HRV parameters were measured in 77 patients(group A)and 41 healthy controls(group B). Long-term linear parameters included standard deviation of normal-to-normal RR intervals(SDNN), standard deviation of all mean 5-minute RR intervals(SDANN), mean of standard deviation of all 5-minute RR intervals(SDNN Index), root mean square successive difference between adjacent RR intervals(RMSSD)and percentage of NN50 in the total number of RR intervals(pNN50). Short-term linear parameters included normalized low frequency(LFn), normalized high frequency(HFn)and low frequency to high frequency(LF/HF). Short-term nonlinear parameters included center-edge ratio(CER), cumulative energy(CE), Lyapunov exponents(LEs), cardiac vagus index(CVI)and cardiac sympathetic index(CSI). The correlation between HRV parameters and the number of 24-hour ventricular premature beats in group A was analysed. Results   Compared with group B, SDNN, SDANN, SDNN Index, RMSSD, pNN50, HFn, LEs and CVI in group A were significantly lower(P<0.05), and LFn, LF/HF, CER, CE and CSI in group A were significantly higher(P<0.05), while both CER and LEs had increasing trends with the worsening of ventricular premature beats(P<0.05). The number of 24-hour ventricular premature beats were positively correlated with CER, CE and CSI(P<0.05), while negatively correlated with LEs and CVI (P<0.05). Conclusion   In patients with essential hypertension and ventricular premature beats, HRV descends significantly and the number of ventricular premature beats increases along with the increased sympathetic tone. Nonlinear HRV parameters are more sensitive in assessing the function of automatic nerve.

    Mutation screening of TFAP2B gene in children with patent ductus arteriosus
    SUN Jin, HAN Bo, WEI Mei-li, YANG Wen-wei, WANG Jie-zhong, LIU Yong-jiao
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  72-75. 
    Abstract ( 1279 )   PDF (1682KB) ( 628 )   Save
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    Objective   To detect TFAP2B gene sequences in PDA children, so as to analyze the relationship between PDA and TFAP2B gene mutation. Methods   A total of 74 children with PDA and a total of 100 healthy children as controls were registered. The peripheral blood was collected and the genome DNA was extracted.All exons and flanking intronic regions of TFAP2B gene were amplified by polymerase chain reaction and the amplions were sequenced by using Sanger sequencing. The required sequences were compared with Genbank and NCBI sequence databases so as to identify whether the mutation or the polymorphism existed.The difference of polymorphic frequency distribution between the PDA children and the controls was tested by Chi-square test.  Results   One single nucleotide polymorphism (c.1-34G→A)of TFAP2B gene was identified in the PDA children and the healthy children. However, the polymorphic frequency distribution was not statistically different from that in the controls(P>0.05).  Conclusion   The single nucleotide polymorphism(c.1-34G→A)of TFAP2B gene may be not related to PDA.

    Status and influencing factors of secondary prevention of ischemic stroke in patients with hypertension
    CAI Yi, LONG Fa-qing, ZENG Chao-sheng, SU Qing-jie, WU Hai-rong, WU Ying-man, LI Peng-xiang, ZHOU Jing-xia,WANG De-sheng, ZHANG Yu-hui
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  76-79. 
    Abstract ( 1034 )   PDF (808KB) ( 449 )   Save
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    Objective   To investigate the status of hypertension preventing in secondary prevention of ischemic stroke  in Hainan province of China and identify the influencing factors. Methods   586 ischemic stroke patients who met the conditions were enrolled in the study. Antihypertensive therapy status of the patients were investigated. Univariate analysis and multivariate logistic regression analysis were performed to find the independent risk factors. Results   Patients’ awareness rate for antihypertensive therapy was 77.1%, treatment rate was 56.0% and control rate was 21.3%. Control rate of the patients who received health education was higher than those who didn’t(P<0.001). Multinomial logistic regression analysis showed that education, health education, type of medical insurance, recommendation of taking medicines  from doctors and history of hypertension were influencing  factors(P<0.05). Conclusion   Patients’  awareness rate, treatment rate and control rate of antihypertensive therapy are low in ischemic stroke patients in Hainan province. Clinical doctors should strengthen health education in ischemic stroke patients and recommend them to take medicines.

    Comparative proteomic study of epileptic foci in  patients with epilepsy
    ZHANG Huan-li1, CUI Ya-zhou2, ZHOU Xiao-yan2, SHANG Wei1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  80-84. 
    Abstract ( 1112 )   PDF (3859KB) ( 628 )   Save
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    Objective   To construct differential express frofilings of brain tissues obtained from epilepsy patients for exploring  the pathogenic mechanism and potential therapeutic targets of epilepsy. Methods   Differentially expressed proteins in epileptic foci of six patients were identified by two-dimensional fluorescence difference gel electrophoresis(2D-DIGE) and matrix-assisted laser desorption/ionizartion time of flight mass spectrometry (MALDI-TOF-MS). Results   15 differentially  expressed proteins which showed at least 1.5 fold difference were detected in the six patients, of which 13 were down-regulated and 2 were up-regulated. Conclusion   Many differentially expressed proteins in epilepsy patients are identified, which may be involved in the pathogenesis of epilepsy.

    Comparative study of endoscopic saphenous vein harvesting applied to different vein calibers
    ZHANG Sen, YANG Chang-yong, ZHANG Gong, SONG Yi
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  85-88. 
    Abstract ( 1317 )   PDF (2208KB) ( 487 )   Save
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    Objective   To compare the influence of saphenous diameters on the operative progress and vessel condition of endoscopic saphenous vein harvesting (ESVH). Methods   The clinical cases of 80 patients who were treated with ESVH from 2011 to 2012 in our hospital were investigated. Patients were divided into two groups by preoperative ultrasound: group A (SV caliber <3mm, n=19), and group B (SV caliber≥3mm, n=61). Total procedure time and EVSH time, SV characteristics such as the total number of side branches, number of ligated branches, number of repaired branches, and total graft length were recorded in the operating room.Light microscopy was used to observe the structure of the vein intima, scanning electron microscopy was used to assess the trauma of the vein intima, and the cell apoptosis was detected by TUNEL assay. Results   Group A had a larger number of side branches (A, 12.9±3.2; B, 9.3±4.1,P<0.05), but graft length [A, (28.2±3.9)cm; B, (27.7±3.1)cm] and ratio of repaired side branches relative to total branches per graft [A, (20.3±9.1)%; B, (18.5±8.0)%] showed no significant inter-group differences(P>0.05). Group A patients required a longer operating time [A, (59.5±12.8)min; B, (41.9±15.4)min], but EVSH time showed no difference(P>0.05).Vascular histological injury was statistically significant(P<0.05),but the cell apoptosis detected by TUNEL assay showed no significant differences(P>0.05). Conclusion   SV with a caliber of <3mm prolong the procedure time and injure the quality of SV in comparison with SV measuring ≥3mm in caliber. So SV with a small caliber is less appropriate for ESVH and is better harvested by a conventional open procedure.

    Relationship between regulatory T cells, IL-35 and aGVHD after allo-HSCT
    SUN Yuan-xin1, QIN Xue-mei1, LIU Xi-min2, WANG Li-zhi2, LIU Chuan-fang1, LIU Lu1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  89-94. 
    Abstract ( 1243 )   PDF (3244KB) ( 740 )   Save
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    Objective   To research the relationship between regulatory T cells (Tregs), IL-35 and acute graft-versus-host disease (aGVHD) in clinical allogeneic bone marrow transplantation (allo-BMT) recipients, and conduct a prospective analysis of peripheral blood Tregs and IL-35 in 23 allo-BMT patients. Methods   The percentage of CD4+CD25+Foxp3+ Tregs in peripheral CD4+T cells was detected by flow cytometry. The relative expression of Foxp3  mRNA transcripts in the mononuclear cells was detected by real time polymerase chain reaction (RT-PCR). And the level of IL-35 in the plasma was got by Enzyme Linked Immunosorbent Assay (ELISA). Results   The percentage of CD4+CD25+Foxp3+ Tregs, the level of Foxp3 mRNA transcripts and the level of IL-35 in the recipients with aGVHD (n=13) were all significantly lower than those in patients without aGVHD (n=10) and the healthy people (n=21) (P<0.05). Tregs decreased linearly with increasing grades of GVHD at onset. Tregs at onset of aGVHD predicted the response to the  aGVHD treatment. Paired analysis in aGVHD patients comparing Tregs at aGVHD onset to prior time points (7-14 days) showed significantly decreased in the ratio of CD4+CD25+Foxp3+ regulatory T cells and the level of Foxp3 mRNA(P<0.05). Conclusion   The ratio of CD4+CD25+Foxp3+ Tregs, Foxp3 mRNA relative transcripts and the level of IL-35 were all decreased in the patients with aGVHD which may have an important prognostic value as a biomarker of acute GVHD.

    Clinical analysis of recombinant human endostatin combined with gemcitabine and  cisplatin therapy in advanced non-small cell lung cancer
    ALIMUJIANG·Stiwaerdi, ZHANG Tao, PATIGULI·Aerxiding, LIU Li
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  95-98. 
    Abstract ( 1067 )   PDF (1726KB) ( 282 )   Save
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    Objective   To investigate the clinical efficacy and adverse reactions of recombinant human endostatin (YH-16) in combination with gemcitabine and cisplatin (GP) in the treatment of advanced non-small cell lung cancer (NSCLC). Methods   80 cases of advanced NSCLC were randomly divided into the observation group (40 cases) and the control group (40 cases). The observation group were treated with YH-16 combined with GP, and the control group accepted GP. After 2 cycles of chemotherapy, the curative effect and adverse reactions were evaluated. The long-term efficacy of the two groups was evaluated during follow-up. Results  The clinical benefit rates in the observation group and the control group were 32.5% and 25.0% respectively, and the difference was statistically significant (P<0.05). But the long-term efficacy and adverse reactions in the two groups were similar (P>0.05). Conclusion   YH-16 in combination with GP in the treatment of advanced NSCLC improves the patient′s curative effect, but does not increase the incidence of adverse reactions.

    CT performance of benign adrenal fat-containing tumor
    WANG Lin-sheng1, ZHANG Li-hong2, CHEN Yue-qin1, SUN Zhan-guo1, WANG Jie-huan1, WANG Yu-hong1, LI Hui1, LI Chuan-fu3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  99-103. 
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    Objective   To investigate the CT performance of adrenal fat-containing tumor. Methods   CT manifestations of 19 cases with adrenal fat-containing tumor confirmed by pathology were retrospectively analyzed. 19 cases were performed with CT plain scan,  including 9 cases with enhanced CT scan. Results   There were 13 cases of myelolipoma, 2 cases of lipoma, 2 cases of teratoma, 1 case of angiomyolipoma and 1 case of lipomatous ganglioneuroma. All cases appeared mixed densities of fat tissues with distinct edges. Among 13 cases of myelomas, 11 cases based on fatty components, and the myeloid components were predominant in 2 cases. There were 3 cases which showed calcification. Enhanced CT showed mild enhancement in the myeloid components of 6 cases. 2 lipoma cases showed uniform fat densities with clear borders. 2 mature teratoma cases containing multiple calcifications, soft-tissues and fat tissues, and the soft-tissue components were mildly enhanced. CT showed that parts of the solid components were markedly enhanced in the angiomyolipoma. The fatty component of the lipomatous ganglioneuroma was predominant, and the soft-tissue component was enhanced gradually. Conclusion   Different types of benign fatty tumors were noted in the adrenal gland. Their CT appearances were some features.

    Study on the learning curve for total laparoscopic hysterectomy
    XIONG Li1, LUO Chun-fang2, ZHOU Jin-tao1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  104-106. 
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    Objective   To explore the learning curve of total laparoscopic hysterectomy(TLH). Methods   85 patients who received TLH were randomly divided into four phases according to their operation time, including 20 cases of Phase 1~3, 25 cases of Phase 4. The operation time and blood loss were compared and analyzed, and the learning curve was fitted with the simple sine curve. Results   The blood loss in Phase 4 was significantly lower than that in Phase 1(P<0.05). The operation time in Phase 3 and Phase 4 was significantly less than that in Phase 1 and Phase 2(P<0.05),while the difference was not significant between Phase 3 and Phase 4(P>0.05). Moving average line was fitted with the simple sine curve Y=sin-π/2, and the inflection point was 41. Conclusion   The learning curve for TLH1 is approximately 40 cases. And the surgeon can achieve a proficient level and stable degree.

    β-catenin expressions in different subtypes of breast cancer and its clinical significance
    LI Shan-shan, SUN Ying, DING Huan, LI Li
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2013, 51(3):  107-110. 
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    Objective   To investigate the expressions of β-catenin in 1different subtypes of breast cancer and its clinical significance. Methods   β-catenin expression was detected in 38 cases of triple negative breast cancer(TNBC)、 131 cases of non-TNBC and 10 cases of normal breast tissues by using tissue microarrays and immunohistochemistry. Results   β-catenin protein was only observed on the cytomembrane of normal breast tissues;However, β-catenin protein was not only observed on the cytomembrane of breast cancer tissues, but also in the cytoplasm and nucleus. Abnomal β-catenin expression was higher in TNBC cases(78.9%) than that in non-TNBC cases(60.3%)(P<0.05). An association was identified between abnormal β-catenin expression and lymph node metastasis, histological grade, TNM stage, Ki67-positive and prognosis(P<0.05). Conclusion   β-catenin may play an important role in breast cancer tumorigenesis and progress, and it could be used as a new indicator for the biological behavior、prognosis and targeted therapy in breast cancer hopefully.