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Table of Content

      
    25 December 2012
    Volume 50 Issue 12
    Articles
    Functional analysis in the intron and 10 kb upstream
    regions of human homeobox gene NKX3.1
    YU Chun-xiao1,2,3, JIN Tong4, JIANG An-li5, ZHAO Jia-jun1,2,3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  13. 
    Abstract ( 1078 )   PDF (2162KB) ( 588 )   Save
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    Objective   To further illustrate regulation mechanisms on the expression of homeobox gene NKX3.1, the intron and the 10kb upstream region of NKX3.1 gene were cloned and their effects on NKX3.1 promoter activity were determined to find the androgen response element and the tissue-specific enhancer. Methods   According to the sequence of NKX3.1 gene in Genbank, a NKX3.1 promoter-luciferase reporter plasmid was constructed by PCR technology. Further, an intron-promoter-luciferase reporter plasmid and 10 fragments-promoter-luciferase reporter plasmids were constructed separately. After recombinated plasmids were transfected into  prostate cancer cell line LNCaP, the effects of the intron or 10 fragments in the upstream of NKX3.1 gene on the promoter activity were tested by luciferase reporter assay. Androgen response region was analyzed by stimulating LNCaP cells with R1881. Prostate-specific enhancer was analyzed by the transfection of plasmids in different cells. Results   The region between -7681~ -6483bp enhanced the activity of NKX3.1 promoter to 2.6 fold, but it was not tissue-specific. The intron and other 9 fragments could not increase the NKX3.1 promoter activity obviously. R1881 didn’t enhance the activities of intron and 10kb upstream region of  NKX3.1.  Conclusion   The intron and 10 fragments of NKX3.1 gene were not androgen-responsible. A functional positive regulatory element existed from -7681~-6483bp, but it was not tissue-specific.

    Expression of negative immunoregulatory gene A20 in human
    hepatocellular carcinoma  and construction of
    its eukaryotic expression vector
    XING Qiao-ling1, HAN Zhen-long2, SONG Yong-hong3, GUO Chun1, ZHU Fa-liang1, ZHANG Li-ning1, SHI Yong-yu1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  19. 
    Abstract ( 1033 )   PDF (2288KB) ( 471 )   Save
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    Objective   To explore the role of negative immunoregulatory gene A20 in human hepatocellular carcinoma(HCC), we investigated the expression of A20 gene in HCC and constructed the A20 eukaryotic expression vector. Methods   The expressions of A20 gene in tumor tissues and tumor-adjacent tissues from 46 HCC patients were detected by immunohistochemistry. The A20 eukaryotic expression vector was constructed and transfected into SMMC-7721 HCC cell lines by the liposome. The mRNA and the protein levels of A20 were detected by the RT-PCR and Western blot methods. Results   A20 expression in tumor tissues was significantly lower than that in tumor-adjacent tissues(P<0.05). A20 expression was closely associated with the tumor differentiation degree and size (P<0.05). The A20 eukaryotic expression vector was successfully constructed which induced the overexpression of A20 in SMMC -7721 cell lines. Conclusion   Negative immunoregulatory gene A20 may be involved in the tumorigenesis of HCC. The A20 eukaryotic expression vector is successfully constructed which can be an important tool for the further study.

    Construction of eukaryotic expression vector for human miR-147a and
    its effects on the proliferation and invasion of lung cancer cells
    YU Yang1, ZHAO Jian2, ZHONG Kai-ze2, LIU Chun-yan1, LI Yi3, SHI Jing3, CHEN Wei-wen1, JIANG An-li1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  25-30. 
    Abstract ( 1073 )   PDF (1873KB) ( 383 )   Save
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    Objective   To construct the recombinant eukaryotic expression vector for human miR-147a and determine its effects on the proliferation and invasion of lung adenocarcinoma cell A549 due to miR-147a expression. Methods   pri-mir-147a sequence was amplified by RT-PCR using total RNA from A549 cells and was inserted into the pSilencer4.1CMV neo expression vector to construct the recombinant pSilencer4.1-147a vector, which was transfected into A549 cells, followed by qRT-PCR and reporter gene assay to detect the expression of miR-147a. MTT assay was utilized to determine the effect of exogenous miR-147a over-expression on the proliferation of A549 cells. Matrigel invasion assay was performed to detect the effect of exogenous miR-147a over-expression on the invasion of A549 cells. Finally,reporter gene assay was used to observe the effects of miR-147a on common tumor signal pathways. Results   The construction  of pSilencer4.1-147a was confirmed to be correct by restriction enzyme digestion and DNA sequencing. qRT-PCR and reporter gene assay displayed significant ectopic expression of miR-147a after the transfection of pSilencer4.1-147a in A549 cells. MTT assay and Matrigel invasion assay showed that miR-147a significantly inhibited the proliferation and invasion ability of A549 cells. Reporter gene assay showed that miR-147a significantly down-regulated relative luciferase activity of pGL4.23-AP1, pGL4.23-ELK1 and pGL4.23-cMYC. Conclusion   The eukaryotic expression vector of human miR-147a is constructed successfully and expressed in A549 cells effectively. The exogenous miR-147a over-expression shows inhibitive effects on the proliferation and invasion of A549 cells, which maybe relevant to the inhibition of EGFR-RAS-MAPK pathway by miR-147a.

    Preparation of 131I-rFliC and its biodistribution
     in breast cancer-bearing mice
    YANG Huan, LIANG Ting, ZHANG Chao, SONG Jing, HAO Jing, HOU Gui-hua
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  31-36. 
    Abstract ( 1009 )   PDF (2871KB) ( 722 )   Save
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    Objective   To investigate the biodistribution and significance of 131I- rFliC in breast cancer-bearing mice. Methods   ① The expression of TLR5 was assayed by the immunofluorescence method,RT-PCR and Western blot. ② rFliC was labeled by 131I and the radiochemical purity,  stability,  cell uptake and efflux of 131I- rFliC were identified. ③  MCF-7 bearing mice were prepared and injected with 131I- rFliC. The biodistribution of 131I- rFliC and tumor targeting were analyzed. Results   ① MCF-7 could express TLR5 at the mRNA and protein levels. The expression of TLR5 was down-regulated by rFliC with a dose-dependent manner. ② The labeling efficiency of 131I-rFliC was 95.19% and the radiochemical purity was 96.46%. 131I-rFliC was more stable in serum and could be absorbed by MCF-7 stably. ③ 24h after injection with 131I-rFliC,  T/M ratio was 7.09, and tumor tissue appeared clearly with the autoradiography. Conclusion   Tumor-targeting of 131I -rFliC is obvious,  and it may become a new molecular probe for the diagnosis and therapy of breast cancer.

    The construction of MHC-Ⅰ intrabody recombinant AAV vector
    WANG Xiao-ping, ZHANG Rui-bin, ZHU Bin, GAO Qing-zhen,JING Yong-sheng, REN Wan-jun, WANG Pu, WANG Qi
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  37. 
    Abstract ( 1138 )   PDF (1458KB) ( 517 )   Save
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    Objective   To construct major histocompatibility complex-I (MHC-Ⅰ) recombinant adeno-associated virus (rAAV) vector. Methods   The gene fragment of endocytoplasmic reticulum stagnation MHC-Ⅰ-intrabody was synthesized followed by sequencing. The intrabody gene fragment was then obtained by restriction enzyme. The intrabody gene fragment was inserted into the EcoR I-BamH I site of the plasmid pSSHG/CMV to construct pSSHG/MHC-Ⅰ. The rAAV viral stock was packaged in 293 cell lines through co-transfecting with the pSSHG/MHC-Ⅰ,  pAAV/Ad and pFG140 instead of adenovirus by calcium phosphate precipitation. Digoxin labeled dot blot hybridization was used to determine the rAAV titer. Results   The recombinant viral stock vector of plasmid pSSHG/MHC-Ⅰ was successfully constructed. The results of dot blot hybridization showed that the rAAV stocks of high titre 6.88×1010 PFU/mL had been obtained. Conclusion   rAAV-MHC-Ⅰ is successfully constructed by molecular cloning and recombination in vitro techniques,which can serve as the foundation of further human cell experiments and gene therapy of transplantation immunity.

    Anti-inflammatory effect of burdock fructo-oligsaccharide on
    lipopolysaccharide-stimulated RAW264.7 cell
    LIU Jing1, PAN Xiao-hua2, SONG Zhen1, CHENG Mei-hua3, WANG Molin1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  41. 
    Abstract ( 1307 )   PDF (2777KB) ( 569 )   Save
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    Objective   To investigate the anti-inflammatory effect of burdock fructo-oligsaccharide (BFOS) on the inflammatory model of RAW264.7 cells. Methods   The inflammation model was constructed by treating RAW264.7 cells with lipopolysaccharide (LPS), and treated with different doses of BFOS. Expressions of Cyclooxygenase (COX-2), Nitric oxide synthases (iNOS), monocyte chemotactic protein-1 (MCP-1) and Interleukin-6(IL-6) were detected by realtime PCR. Expressions of MCP-1 and IL-6 were determined by ELISA before and after the treatment with BFOS. Expressions of COX-2, iNOS, phosphorylated IκB, NF-κBp65, extracellular signal-regulated (Erk), P38 and c-Jun Nterminal kinase (JNK) were detected by Western blot. Results   The expression levels of iNOS,COX-2,MCP-1 and IL-6 of RAW264.7 cells stimulated by LPS increased, and BFOS significantly decreased them . BFOS decreased the expressions of p-IκB,p-NF-κBp65,p-Erk,p-P38,and p-JNK compared to the control. Conclusion   BFOS may exert anti-inflammatory effect on RAW264.7 cells stimulated by LPS.

    Model construction of transplanting rat embryonic skin
    precursor complex in nude mice
    LI Chuan, CHENG Yan-na, JIANG Du-yin
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  47-51. 
    Abstract ( 888 )   PDF (3225KB) ( 379 )   Save
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    Objective   To screen the optimal pregnancy time window of transplanting rat embryonic skin precursor tissue in nude mice, and explore a new method for studying and intervening embryonic skin and appendage development. Methods   Rat embryonic skin precursor complexes(ESPCs) of different gestational ages(E14.5d, E16.5d and E18.5d) were acquired with little subcutaneous tissue, then they were implanted under dorsal flap of nude mice. 7 days after transplantation, the flap was opened and living ESPCs was exposed. The histological characteristics of fetal skin and development of appendages were observed. Results   E16.5d and E18.5d grafts survived successfully and continued developing, and finally generated intact skin structure with skin appendages. E14.5d grafts generated into teratoid neoplasm. Conclusion   With the methods of implanting under flap, ESPCs can successfully survive, and E16.5d is the optimal time for transplanting ESPCs.

    Effect of hemimastication on the mitochondria of rat masseter muscle
    WANG Xiao-fei1, DING Ting-ting1, LIU Zhao1, JI Ping1,2, LIN Xue-fen1, QI Dong1, FENG Dan-dan1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  52. 
    Abstract ( 1030 )   PDF (1626KB) ( 408 )   Save
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    Objective   To explore the effect of hemimastication on the mitochondria of rat masseter muscle and to study the mechanism of masseter muscle dysfunction induced by hemimastication. Methods   Mitochondrial morphology and number changes were observed by electron microscope, and the mitochondrial damage index was measured; Ca2+ content was measured with atomic absorption spectrometry; The mitochondrial permeability transition pore(MPTP) opening was detected by UV spectrophotometry. Results   The mitochondrial damage index score of the extraction side was lower than that of the non-extraction side, and significantly lower than that of the control group(P<0.01). The 4-week group had the lowest score.Compared with those of the control groups, Ca2+ contents in the experimental groups had the higher level except the 8week group (P<0.05). The opening of mitochondrial permeability transition pore had a higher level on the extraction side than that on the non-extraction side and the control group(P<0.05). The 6-week group had the highest level. The opening of mitochondrial permeability transition pore was negatively correlated to mitochondrial damage index (r=-0.74,P<0.05). Conclusion   The signal way of the mitochondrial permeability transition pore opening can be activated by high concentration of calcium, which may cause the damage of mitochondria and apoptosis.This pathway may play an important role in the mechanism that hemimastication results in masticatory muscles dysfunction.

    Expression of HSP90β in gastric adenocarcinoma and its      
    clinical pathological significance
    XU Ling1, LIU Wan2, SONG Yan-yan1, SUN Li-dan1, LI Xia3, ZHANG Yuan3, ZHANG Jian-ping1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  56. 
    Abstract ( 1016 )   PDF (1634KB) ( 383 )   Save
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    Objective   To explore the expression of heat shock protein 90β (HSP90β) in gastric adenocarcinoma and its correlation with proliferation and clinicopathological parameters of the tumor cells, and to analyze the relationship of HSP90β with the patients′ prognosis. Methods   Use immunohistochemistry PV-9000 two-step method to detect the expression of HSP90β and Ki-67 in 60 cases of gastric adenocarcinoma and 20 cases of gastritis,  the χ2 test was used to analyze the relationship between clinicopathological parameters,  the Φ coefficient was used to discuss the correlation between the HSP90β and Ki-67,  and survival analysis was performed on 60 patients with gastric cancer. Results   Expressions of HSP90β and Ki-67 in gastric adenocarcinoma were higher than those in gastritis tissues (P<0.05),  and the positive rate in poorly differentiated gastric adenocarcinoma was much higher than that in well and moderately differentiated gastric adenocarcinoma (P<0.05);there was a positive correlation of expression between HSP90β and Ki-67 in gastric adenocarcinoma(Φ=0.368,  P<0.05).The survival period of patients with HSP90β positive expression was less than that of patients with negative expression. Conclusion   HSP90β is significantly up-regulated in gastric carcinoma. Its expression is positively correlated to Ki-67 and heralded a poor prognosis.

    Expression of activation induced cytidine deaminase and its
    relationship with helicobacter pylori in the development
    of intestinal-type gastric cancer
    ZHANG Qi1,2, LI Yan-qing1, ZHANG Cui-ping2, ZHAO Kun2, SUN Xiang-hong3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  61. 
    Abstract ( 1140 )   PDF (2222KB) ( 323 )   Save
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    Objective   To investigate the expression of activation induced cytidine deaminase (AID) in the development of intestinal-type gastric cancer(IGC) and the relationship with H.pylori infection, and to investigate the role of AID and H.pylori in the carcinogenesis. Methods   The expressions of AID in the chronic superficial gastritis (CSG),chronic atrophic gastritis (CAG) with intestinal metaplasia (IM) and intestinal-type gastric cancer(IGC) were detected with the immunohistochemical method, and H.pylori was detected. The relationship between AID and CDX2 was analyzed. Results   The expression rates of AID in the H.pylori- positive groups were higher compared with the H.pylori-negative groups(P<0.05). The expression rates of AID in CAG with IM and IGC were higher compared with CSG(P<0.05). The expression of AID was positively correlated to the expressions of CDX2 in CAG with IM and IGC(r=0.310, P<0.05; r=0.324, P<0.05). Conclusion   H.pylori may induce abnormal expression of AID in the gastric epithelia and AID may contribute to the development of IGC.

     

    Relations between serum leptin and leptin receptor Gln223Arg
    polymorphism with gastro-esophageal reflux disease
    MA Xing-bin1, CHE Xiao-wen2, WU Shuo1, JIN Yun-ling3, ZHANG Nan1,
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  65. 
    Abstract ( 1009 )   PDF (1019KB) ( 445 )   Save
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    Objective   To investigate the relations between serum leptin and leptin receptor (LEPR) Gln223Arg polymorphism with gastro-esophageal reflux disease(GERD). Methods   62 patients with GERD were selected according to reflux disease questionnaire and gastroscope inspection and divided into the reflux esophagitis (RE) group and nonerosive reflux disease (NERD) group. In addition, 63 healthy subjects were selected as the control group. The serum leptin levels were determined by ELISA, and LEPR Gln223Arg polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphisms assay. Results   The levels of leptin in RE group and NERD group were both significantly higher than in control group (P<0.05); the degree of inflammation in RE group was positively correlated with leptin level. GA+AA genotype and A allele frequency of LEPR gene in RE group and NERD group were significantly higher than those in control group (P<0.05). The leptin in patients with GA+AA genotype was significantly higher than that in patients with GG genotype (P<0.05). Conclusion   The serum leptin and the LEPR Gln223Arg polymorphism are associated with GERD.

    Clinical research on treatment of liver decompensation of primary
    liver cancer by the method of Yiqibushenquyu
    XU Xiao-mei1, YU Jiao1, LI Guo-hua1, JIANG Yan2, JIANG Zhao-quan3, ZHANG Ping3, CHENG Xiao-yun1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  70. 
    Abstract ( 892 )   PDF (937KB) ( 302 )   Save
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    Objective   To observe the clinical efficacy and the safety of Yiqibushenquyu for liver decompensation of primary liver cancer. Methods   60 cases were randomly divided into experimental group (30 cases) and control group (30 cases). Both groups were given conventional treatment of hepatoprotective anti-virus and symptomatic treatment. Besides the conventional treatment, the experimental group was also given the Chinese medicine of Yiqibushenquyu. Results   The total efficacy rate was 60.00% in the experimental group and 33.33% in the control group. After the treatment, patients survived for 19.00 months on average in the experimental group and 9.13 months in the control group. More satisfied efficacy was found in modifying fatigue, increasing albumin, lowering bilirubin, and improving AFP negative conversion in the experimental group. All differences in the above indicators were significant between the two groups. Conclusion   The method of Yiqibushenquyu can reduce patients’ pain, improve liver function, and prolong patients’ survival.

    Preliminary study of lung cancer types with gemstone spectral CT imaging
    LI Ming-ying1, WANG Guang-li2, CUI Ding-ye2, ZHANG Cheng-qi2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  73. 
    Abstract ( 1135 )   PDF (1396KB) ( 527 )   Save
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    Objective   To investigate the preliminary application of spectral CT imaging on different types of lung cancer. Methods   60 patients who were diagnosed lung cancer at first time were scanned in gemstone spectral imaging(GSI) mode by using dual energy CT. Iodine concentration, water concentration, spectrum energy curve and CT values at 40 keV in the region of interest of the mass were measured with GSI post-processing software to analysis the spectral feature of different types of lung cancer. Results   Significant differences in iodine concentration, spectrum energy curve and CT values at 40keV existed between lung squamous cell carcinoma and lung adenocarcinoma(P<0.05), while water concentration were found no significant difference between them(P>0.05). Conclusion   GSI technique may have important value in lung cancer classification by providing qualitative and quantitative informations of tumors.

    Therapy effect of 5-FU palliative implanted through puncturing along
    cardiac tangent direction in 81 patients with malignant pericardial effusion
    LI Ping, LIANG Xiao-hong, JI Yong-li, DONG Liang, GAO Yang, CUI Wen-Chao
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  77. 
    Abstract ( 935 )   PDF (8040KB) ( 442 )   Save
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    Objective   To investigate the clinical value of the 5-FU sustained-release particles implanted through puncturing along the cardiac tangent direction for malignant pericardial effusion. Methods   81 patients with malignant pericardial effusion were treated by pericardiocentesis guided by ultrasound and 5-FU palliative particles were implanted into the pericardial cavity. The puncturing path was along the cardiac tangent direction. Ultrasound examinations were used every week after the treatment. The efficacy was evaluated four weeks after the treatment. The 45 patients treated by pericardial catheter drainage and intracavitary chemotherapy at the same period were served as controls. Results   The puncturing success rate was 100% in the 81 cases. Ultrasound reviews showed that complete remission occurred in 71,partial remission in 8, and no response in 2 cases four weeks after the treatment. The total remission rate was 97.53%, which was significantly higher than that in the control group(77.78%, P<0.01). Conclusion   It is a new ideal method to treat malignant pericardial effusion to implant 5-FU sustained-release particles through puncturing along the cardiac tangent direction for its high safety and efficacy and less adverse reactions.

    Effects of pregnancy and childbirth on female pelvic  support floor
    WANG Jing-chen1,2, YUAN Miao2, LIU Guang-li2, FENG Hao2, RONG Feng-nian2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  82-86. 
    Abstract ( 1088 )   PDF (926KB) ( 375 )   Save
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    Objective   To explore the effects of pregnancy and childbirth on  postpartum early pelvic floor function and the occurrence of pelvic floor dysfunction  disease(PFD). Methods   A total of 568 full-term primipara, who had the follow-up at 6~8 weeks after delivery were included. Test pelvic floor muscle strength, fatigue strength and pelvic floor pressure by digital vaginal palpation and equipment to explore pelvic floor function; general questionnaires and pelvic organ prolapse quantitive examination (POPQ) scoring were used to know early postpartum PFD and influencing factors. Results   The normal rate of typeⅠand typeⅡ-muscle fiber strength was 29.40% and 33.10%; the normal rate of fatigue strength was 52.82% and 89.08%; the normal rate of pelvic floor pressure was 72.54%. There were no significant difference between the different delivery groups(P>0.05). The incidence of stress urinary incontinence (SUI) in pregnancy was 27.82 %. There was no significant difference between the different delivery groups(P>0.05).The incontinence of SUI, vaginal anterior and posterior prolapse , uterine prolapse at 6~8 weeks after delivery was respectively 9.86%, 79.40%, 21.13%, 12.50% .And the incontinence was vaginal delivery group>cesarean section group, with significant difference between them(P<0.05). It showed that vaginal delivery, lateral episiotomy and newborn birth weight(≥4000g)were risk factors of postpartum SUI and POP.  SUI during pregnancy was also the risk factor of postpartum SUI. Conclusion   Pregnancy and childbirth make pelvic floor function decline and postpartum SUI and POP appear, which should be paid attention to early.

    Expression of steroidogenic enzyme in subclinical Cushing’s
    syndrome due to adrenal adenomas
    YANG Xue-cheng1, SUN Rui-xia2, FAN Liang-liang3, WANG Kai-ping2, JIAO Wen-juan2, CAO Cai-xia2, GAO Yan-yan2, SUN Li-jiang1, WANG Xin-sheng1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  87. 
    Abstract ( 1067 )   PDF (2023KB) ( 391 )   Save
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    Objective   To determine whether steroidogenic enzyme expression is associated with the steroid secretory pattern of subclinical Cushing’s syndrome (SCS) by investigating the expression of CYP17 and CYP11B1 in SCS adenomas, normal adrenal cortices (NA), non-functional adrenal adenomas (NFA) and cortisol-producing adenomas (CPA). Methods   Total RNA and protein were extracted from 20 CPA, 6 SCS, 15 NFA, and 8 NA. Real-time quantitative polymerase chain reaction (PCR), western blotting and immunohistochemistry analysis were performed to determine the mRNA and protein expression of CYP17 and CYP11B1 in different tissues. Expressions of CYP17 and CYP11B1 in the adrenocortical tumors were compared in NA. Results   Expressions of both CYP11B1 and CYP17 mRNA and protein were detected in all samples tested. Expressions of CYP11B1 mRNA and protein were significantly higher in CPA group than the others, and were slightly higher in SCS samples than that in NA and NFA samples (P<0.05). Expressions of CYP17 mRNA and protein in different tissues were similar to that of CYP11B1. They showed positive correlations with plasma cortisol and negative correlations with adrenocorticotrophin levels (P<0.05). Conclusion   CYP11B1 and CYP17 were over-expressed in subclinical CPA,and their over-expressions were caused by the increased production of cortisol in CPA.

    The effect of methyltransferase inhibitor on the expression of
    XAF1 mRNA in prostate cancer
    GUO Xiao-yu1, JIANG Yan2, XING Zhao-quan3, GUO Zhao-xin3, FANG Zhi-qing3, LIU Zhao-xu1,3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  94. 
    Abstract ( 924 )   PDF (1179KB) ( 388 )   Save
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    Objective   To investigate the expression of XAF1 mRNA and the effect of DNA methylation inhibitor on its expression in prostate cancer.  Methods   Culture prostate cancer cell lines LNCaP, DU145, PC3 cells and renal cell carcinoma-derived A498 cells.Extract normal peripheral blood mononuclear cells (PBMCs). A498 cells and PBMCs were used as positive controls. Collect 10 clinical cases of prostate cancer and neighboring prostate tissue as control. RT-PCR was performed in order to detect the XAF1 mRNA experssion in LNCaP, DU145, PC3, A498 and PBMCs. Methylation-specific PCR(MSP) was used to detect the XAF1 experssion in LNCaP and DU145 cells before and after they were treated with DNA methylation inhibitor (5′-aza).   Results   LNCaP and DU145 prostate cancer cell lines expressed a shorter form of XAF1 transcripts than the A498 cells, whereas PC3 cells exhibited a complete silence of the XAF1 gene.Yet,after being treated with 5′-aza, LNCaP and DU145 expressed the full length of XAF1 mRNA. Conclusion   Low expression of XAF1 mRNA is found in LNCaP and DU14 cells. No expression is found in PC3 cells.DNA methylation inhibitor can induce the full-length of XAF1 mRNA expression in prostate cancer cells.

    Effect of stiring microcarrier system culturing on apoptosis regulation
    of adult degenerative nucleus pulposus cells
    NING Bin1, GONG Wei-ming1, LI Ling-mei2, GUO Jie1, XIA Qi-gan1, NING Hong1, JIA Tang-hong1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  98. 
    Abstract ( 915 )   PDF (1533KB) ( 463 )   Save
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    Objective   To investigate the effects of nucleus pulposus cells apoptosis on degenerated intervertebral discs by using cell amplification with microcarrier system and studying cell apoptosis. Methods   After primary culture of the degenerative nucleus pulposus tissues by operation, nucleus pulposus cells were randomly separated into two groups. Of the two groups, one was cultured with monolayer cluture, another with microcarrier system. In order to detect apoptosis of different stages, Annexin-V-PI staining was used for early stage apoptosis, DeadEndTM staining for late stage apoptosis, immunohistochemical staining for Bcl2 protein. Results   Annexin-V-PI staining demonstrated that apoptosis rate of nucleus pulposus cultured with microcarrier system was lower in stable phase (P<0.05). However, there was no significant difference between the two different culture methods in logarithmic growth phase (P>0.05).  The data of DeadEndTM staining and immunohistochemical staining showed that apoptosis rate of nucleus pulposus cultured with microcarrier system was lower in both phases (P<0.05). Conclusion   Microcarrier system, which can efficiently reduce the apoptosis rate of nucleus pulposus cells, is a promising culture method applied in tissue engineering of intervertebral discs.

    Clinical research of atlantoaxial transarticular screw fixation for odontoid
    fractures with computer navigation assisted system
    ZUO Can-hui, WANG Lu-bo, XU Shi-hong, DONG Jin-lei, ZHOU Dong-sheng
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  103. 
    Abstract ( 969 )   PDF (1513KB) ( 326 )   Save
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    Objective   To assess the clinical value of atlantoaxial transarticular screw fixation for odontoid fractures with computer navigation assisted system through comparing the clinical effects of atlantoaxial transarticular screw fixation by computer navigation assisted system (navigation group) with those of atlantoaxial transarticular screw fixation by C-arm fluoroscopy(historic control group). Methods   We retrospectively reviewed twentiy-eight patients diagnosed with odontoid fractures who underwent posterior approach atlantoaxial fusion by transarticular screw fixation. The navigation group included 13 patients and the other 15 patients were in the historic control group. Operative time,  fluoroscopy time,  blood loss and postoperative complications were compared between the two groups. We evaluated the screw accuracy through postoperative radiology examination. Results   Totally 55 atlantoaxial transarticular screws were placed in the twentiy-eight patients. The mean operative time was 154.9 min in the navigation group compared with 182.2 min in the historic control group.There was statistical difference betwe-n the two groups(P<0.01).The mean fluoroscopy time in the navigation group was 41.3s  compared to 64.3s in the historic control group (P<0.01). Postoperative radiographs showed all the screws were placed satisfactorily in the navigation group. The position of six screws in the historic control group was demonstrated suboptimal. Conclusion   The computer navigation assisted system enhanced safety and accuracy of atlantoaxial transarticular screw fixation,  decreased patients and surgeons’ radiation exposure and reduced operation time. This technology is an effective method for the treatment of odontoid fractures.

    Mutation analysis of SQSTM1 and TNFRSF11A genes in
    a Chinese family with Paget disease of bone
    SUN Ya-fang1, WANG Yin-chang2, WANG Lai-cheng3, JIAO Yu-lian3, CUI Bin3, XIA Yu3, LU Bing-ru3, ZHAO Yue-ran1,3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  107-113. 
    Abstract ( 1134 )   PDF (2619KB) ( 562 )   Save
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    Objective   To investigate whether mutation in SQSTM1 or TNFRSF11A genes was the genetic cause of Paget’s disease of bone(PDB) in a Chinese pedigree.  Methods   One Chinese family with Paget’s disease of bone was examined. Genomic DNA was extracted from the blood samples of the patients and unaffected members of the pedigree. Polymerase chain reaction(PCR) was performed to amplify all exons of the SQSTM1 gene and TNFRSF11A gene and the PCR products were directly sequenced to detect the mutation. The sequence results were compared to the normal sequence of GenBank.  Results   No mutation but 14 previously reported single nucleotide polymorphisms(SNP) were identified in the two genes. Conclusion   The PDB in this pedigree is not associated with any of the SNPs. PDB in this pedigree may be caused by genes other than SQSTM1 and TNFRSF11A.

    Role of the Th17 cells in oral lichen planus
    YIN Meng, QI Xiang-min, YAN Shi-guo, WEI Mei-rong, CHEN Dan, SONG Jing, ZHANG Da
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  114. 
    Abstract ( 1118 )   PDF (914KB) ( 386 )   Save
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    Objective   To detect the expressions of IL-17mRNA and IL-23mRNA in epithelium and lamina propria of oral mucosa and investigate the role of Th17 cells on oral lichen planus(OLP). Methods   Oral mucosa from 10 cases with OLP were taken as research objects and 6 healthy volunteers were the control. Real-time RT-PCR was used to detect the expressions of IL-17 and IL-23 mRNA of the normal and OLP oral mucosa. Results   The expression levels of IL-17mRNA and IL-23mRNA in both epithelium and lamina propria of the OLP group were significantly higher compared with the normal oral mucosa group (P<0.05) . In the OLP group, the expression level of IL-17mRNA in lamina propria was significantly higher than that in the epithelium (P<0.05). However, the expression levels of IL-23 mRNA showed no significant difference between the lamina propria and epithelium(P>0.05). Conclusion   The increase of cytokine mRNA expressions related to the function of Th17 cells may be correlated to the pathogenesis of OLP.

    Clinical observation of the inlays made of different materials in
     restoring the molar defect
     MI Yong1, SUN Lan-ying2, YIN Jing1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  118-121. 
    Abstract ( 940 )   PDF (904KB) ( 476 )   Save
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    Objective   To evaluate and compare the clinical performance of Au alloy inlays, IPS EmpressⅡceramic inlays, Ceramage inlays and resin restoration in restoring the molar defect. Methods   45 Au alloy inlays, 48 Ceramage inlays, 44 IPS EmpressⅡceramic inlays and 52 composite resin were used to restore 189 molars from 138 patients respectively. All restorations were evaluated after half a year according to modified USPHS parameters. Results   There was no significant difference in the color match, marginal sealability, secondary carries and food embed between the three groups (P>0.05). The Au alloy inlays and Ceramage inlays showed a significantly better performance in resisting fracture than the ceramic inlays (P<0.05). There was a significant difference in the gingival response and food embed between the resin fillings and the inlays (P<0.05). Conclusion   It is an intriguing method using inlays to restore the molar defect, which is superior to resin restoration in the aspects of restoring proximal relationship and maintaining periodontal health.

    Knowledge-attitude-practice analysis of rural residents on
    essential medicine system in Shandong province
    LOU Peng-yu, DAI Hai-yan, YI Li
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  122-125. 
    Abstract ( 881 )   PDF (900KB) ( 381 )   Save
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    Objective   To conduct a survey and analyze rural residents’ knowledge-attitude-practice about the essential medicine system in Shandong province of China. Methods   Multi-stage stratified and random cluster sampling was employed for questionnaires from 862 rural residents. Results   The validity rate of all questionnaires was 96.52%. 31.25% of the rural residents knew the system, 79.23% thought it was good, and 40.38% preferred to use essential medicines. The rural residents mainly obtained the knowledge about the essential medicine system from television (58.46%), free publicity materials (43.46%) and doctors (35.00%). Conclusion   The rural residents have low awareness of the essential medicine system, so it is necessary to adjust propaganda ways, enhance the communication with stakeholders, and regulate stakeholders’ behavior.

    Measurement and evaluation of personal radiation dose in
    cyclotron center and PET-CT department
    ZHANG Wei1,2, LI Shi-xue1, LI Lian-bo2, YU Xi-rong2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2012, 50(12):  126-129. 
    Abstract ( 1362 )   PDF (903KB) ( 485 )   Save
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    Objective   To measure and evaluate the personal radiation dose for medical staff in cyclotron center and PET-CT department and to offer the reference data for clinical radiation protection. Methods   γ ray radiation dose rate was analyzed in 18 medical staff members during each medical procedure by using X and γ radiation detector. The estimation of annual exposure dose of 18 medical staff at different working site was based on the caculation of doserate(d), work time(t) and workload(w). Results   When the workloads were 1000, 4000, 7000 and 10000 per year, the annual doses per procedure were: 0.27 mSv for quality control; 0.02 to 0.20 mSv for dispensation; 0.06 to 0.58 mSv for carrying the compound; 2.49 to 24.9 mSv for injection and 1.19 to 11.91 mSv for setting the patients. Conclusion   The staff receives higher radiation dose during dispensation and injection. When the workload is 4000 and 10000 per year, the dose received by hands and body would respectively exceed the national radiation safety standards (GBSS) on occupational annual dose limits.