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Table of Content

      
    16 November 2010
    Volume 48 Issue 11
    Articles
    Effect of metformin on type 1 angiotensin II receptor expression in endothelial cells
    ZHANG Xu1, YU Wen2, WANG Xiao-yong1, GUAN Qing-bo1, ZHAO Jia-Jun1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  1. 
    Abstract ( 1275 )   PDF (862KB) ( 394 )   Save
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    Objective   To observe the effect of metformin on type 1 angiotensin II receptor (AT1R)expression in endothelial cells and investigate its mechanism. Methods     AT1R, inhibitor protein κB (I-κB)and AMP-activated protein kinase α(AMPKα ) protein expressions in endothelial cells were determined by Western blot, and nuclear factor-κB (NF-κB) activation in endothelial cells was observed by con-focal laser scanning microscopy. The lactate dehydrogenase (LDH) activation in the culture medium was detected by colorimetric assay. Also, the roles of NFκB and AMPKα in the effect of metformin on AT1R were determined by RNA interference technology. Results    I-κB phosphorylation and AT1R protein expression induced by tumor necrosis factor α (TNFα) in endothelial cells were significantly reduced by metformin, and  LDH activity in the endothelial cell culture medium was significantly decreased (P<0.01). The inhibitory effect of metformin on AT1R expression in endothelial cells was significantly reversed by AMPKαSiRNA (P<0.01). Conclusion    NF-κB activation and AT1R expression can be inhibited by the intervention of metformin, and activation of the AMPKα pathway plays an important role in the beneficial effect of metformin on cardiovascular endothelial cells.

    ROC analysis on the screening effect of the type 2 diabetes risk score  and its covariate factors
    WANG Xiao-yong1, LI Shi-xue2, SUN Xiu-bin2, QIAO Qing3, PANG Zeng-chang4, ZHANG Xu1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  7-11. 
    Abstract ( 1040 )   PDF (548KB) ( 330 )   Save
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    Objective      To evaluate the effect of the type 2 diabetes risk score (Score) on screening for asymptomatic diabetes and its covariate factors, such as age and gender, by establishing a receiver operating characteristic (ROC) curve model.  Methods    The data from a cross-sectional survey of 4070 subjects who underwent oral glucose tolerance tests (OGTT) for type 2 diabetes screening in the Qingdao Diabetes Prevention Program in 2006 were analyzed. All the subjects filled out the Score designed for the program at the same time. Sensitivity, specificity, and the area under the ROC curve (AUC) of the Score for asymptomatic diabetes screening was calculated. Then the Score was transformed into an ordered variable, and it was used as the dependent variable, and developing diabetes or not, age and/or gender and their interaction as the independent variables to fit an ordered logistic regression model. The AUC was calculated through the model and the influence of the covariates, age or/and gender, on the screening effect of the Score was evaluated.  Results    Sensitivity, specificity and AUC of the Score for asymptomatic diabetes screening were 0.62, 0.62 and 0.67, respectively. The influence of age on the screening effect of the Score was greater in the young age group than that in the old age group, and the influence of gender was greater in women than in men. So the Score had a better effect on screening for asymptomatic diabetes in the young women group.  Conclusion     The Score is a simple and effective tool for screening for asymptomatic type 2 diabetes. In order to achieve a better screening effect,  covariate factors, age and gender should be considered, when developing and conducting the diabetes risk score for population screening.

    Metabolic characteristics and insulin resistance in elderly patients with different thresholds for impaired fasting glucose
    FU Fang-ming1,2, DONG Xiao-lin1,2, WANG Shao-lian1,2, CHEN Li1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  12-15. 
    Abstract ( 1151 )   PDF (358KB) ( 486 )   Save
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    Objective    To investigate metabolic characteristics, insulin resistance (IR) and islet B-cell function in elderly subjects with different thresholds for impaired fasting glucose(IFG). Methods    A cohort of 830 nondiabetic subjects older than 60 years with fasting plasma glucose(FPG)<7.0mmol/L were divided into three groups:the normal glucose tolerance(NGT) group(n=510) with FPG<5.6mmol/L, the IFG1group(n=167) with 5.6mmol/L ≤FPG<6.1mmol/L, and the IFG2 group(n=153) with 6.1 mmol/L ≤FPG<7.0mmol/L.  Height, weight, blood pressure, waist circumference and hip circumference were measured, and the body mass index(BMI) and waist hip ratio(WHR) were calculated. Triglyceride(TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol(LDL -C), FPG and 2h post-75g oral glucose tolerance test(OGTT) plasma glucose(2hPG) were determined. Fasting insulin(FINS) and 2h post-OGTT insulin(PINS)were measured by radioimmunoassay.The homeostasis model assessment (HOMA) was applied to assess the status of insulin resistance (HOMA-IR) and islet B-cell function (HOMA-B). Associations of IFG1 and IFG2 with risks of post-challenge glucose intolerance and metabolic syndrome (MS) were tested by logistic regression analysis with adjustment for age and gender. Results     Compared with the NGT group, age, WHR, BMI, blood pressure, TC, LDL-C, TG, FPG and 2hPG were elevated in the IFG1 and IFG2 groups(P<0.05). Subjects with IFG2 had  significantly higher 2hPG than those with IFG1(P<0.05). After adjustment for age, sex and BMI, HOMA-IR increased in subjects with IFG(P<0.05), and subjects with IFG2 had higher HOMA-IR(P<0.05). Compared with subjects with NGT, subjects with IFG had lower HOMA-B, while there was no significant difference between IFG1 and IFG2(P>0.05). In logistic regression analysis with adjustment for age and gender, IFG2 was associated with higher risks of post-challenge glucose intolerance and MS compared with IFG1. Conclusions    Insulin resistance and B cell dysfuction are present in elderly subjects with different thresholds for IFG. IFG2 is associated with more serious metabolic disorders and insulin resistance, and it is much more likely to have risks of post-challenge glucose intolerance and MS.

    Alterations of first-phase secretion function of islet alpha and beta cells in subjects with normal glucose tolerance and family history of diabetes
    HE Jun1, QIAO Zong-xing2, SUN Xiao-hui3, WANG Yan-gang3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  16-19. 
    Abstract ( 1085 )   PDF (377KB) ( 377 )   Save
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    Objective     To explore the alterations and their implications of first-phase secretion function of islet alpha and beta cell in subjects with normal glucose tolerance, and, family history of diabetes. Methods     40 relatives, with normal glucose tolerant and family history of type 2 diabetes, were enrolled in the observation group (FH+), while 55 healthy volunteers were in the control group (FH-). Fasting blood-glucose(FBG) and 2-hour postprandial sugar(P2hBG) were determined by enzyme technique. Free fatty acid(FAA) was detected by improved colorimetry. Insulin was detected by chemoiluminesence immunoassay, while the ABC-ELISA technique was applied to determine glucagon on an empty stomach and at 2, 4, 6min after L-ARG stimulation. Results     ① The peak value of insulin and glucagon secretion were at 2 minutes, and began to decrease at 4 minutes in the two groups. ② Compared with the FH- group,P2hBG, FINS, HOMAIR and FFA were significantly higher (all P<0.05) , and the peak factor of insulin was obviously lower in the  FH+ group (P<0.05). There were no differernces of FGC and the peak factor of glucagon (P>0.05) between the two groups. ③Correlation analysis showed the peak factor of insulin had positives collinearity with P2hBG and FFA due to that correlation coefficient r  were 0.274 and 0.356, respectively(P<0.05). Conclusions    In genetics, type 2 diabetes mellitus results from the exhaustion of function of beta cells′ firstphase secretion. The peak factor of insulin may be an important predictor for the occurrence and development of type 2 diabetes.

    Role of Tribble 3 on the production of collagen type Ⅰby cardiac fibroblasts induced by high glucose
    TANG Meng-xiong1,2, L Rui-juan1,2, ZHONG Ming2,3, ZHANG Yun2,3, ZHANG Wei2,3, CHEN Yu-guo1,2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  20. 
    Abstract ( 995 )   PDF (370KB) ( 487 )   Save
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    Objective    To investigate the role of Tribble 3(TRIB3) on the production of collagen type Ⅰby cardiac fibroblasts induced by high glucose. Methods    Rat cardiac fibroblasts were cultured in vitro with different concentrations of glucose. mRNA expressions of collagen type Ⅰand TRIB3 were measured by real time-PCR, and protein level of collagen type Ⅰwas determined by enzyme-linked immunosorbent assay. siRNA targeting TRIB3 was transfected into cardiac fibroblasts, and then the production of pro-collagen type Ⅰby cardiac fibroblasts induced by high glucose was detected. Results     Both mRNA and protein expressions of collagen type Ⅰwere increased after being induced by high glucose(P<0.05), and mRNA expression of TRIB3 was also increased(P<0.05). After inhibiting expression of TRIB3 by siRNA, the increased mRNA and protein levels of collagen type Ⅰinduced by high glucose were reversed(P<0.05). Conclusion     TRIB3 is involved in the production of collagen type Ⅰby cardiac fibroblast induced by high glucose.

    Alterations of immunogenicity in the differentiation from human bone marrow derived mesenchymal stem cells to chondrocytes
    GU Yan-zheng1,3, CAI Yan2, NI Li2, HUANG Chen2, YANG Hui-lin2,3, ZHANG Xue-guang1,3, SHI Qin2,3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  24-28. 
    Abstract ( 1252 )   PDF (1597KB) ( 474 )   Save
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    Objective   To investigate expressions of co-stimulatory molecules and their effects on alterations of immunogenicity on human bone marrow derived mesenchymal stem cells and differentiated chondrocytes. Methods    Human bone marrow mesenchymal stem cells (BMSCs) were isolated by density gradient centrifugation and confirmed by an inverted phase contrast microscope and flow cytometry (FCM). BMSCs were induced to differentiate into chondrocytes. Expressions of co-stimulatory molecules were quantified by FCM. T cells were incubated respectively with undifferentiated BMSCs and chondrocytes, while the 3H-TdR incorporation method was carried out to evaluate T cell proliferation. Results     BMSCs did not express co-stimulatory molecules CD28, CD80, CD83 and CD86,but  inhibited the proliferation of T cells. Expressions of CD28, CD80, CD83, and CD86 in hBMSCs and differentiated chondrocytes were up-regulated and proliferation of T cells was promoted. Conclusion     BMSCs can inhibit proliferation of T cells in vitro by increasing immunogenicity of BMSC-differentiated chondrocytes.

    Construction of the recombinant plasmid pEGFP-N1-ZIP10 and its effect  on expressions of other zinc transporters in breast cancer cells
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  29-32. 
    Abstract ( 1064 )   PDF (842KB) ( 258 )   Save
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    Objective    To construct the pEGFP-N1-ZIP10 expression vector and observe its expression in human breast cancer MCF-7 and MDA-MB-231 cell lines, and also to detect expressions of other zinc transporters when ZIP10 is over-expressed. Methods    The target sequence of ZIP10 was obtained and amplified from human blood by RT-PCR. Then, the cDNA segment was cloned into eukaryote plasmid pEGFP-N1. The pEGFP-N1-ZIP10 was identified by restriction enzyme digestion and checked by DNA sequence analysis. MCF-7 and MDA-MB-231 cells were transiently transfected, and expressions of ZIP10 and other zinc transporters were detected by RT-PCR. Results    Identification of pEGFP-N1-ZIP10 by enzyme digestion and PCR showed that the length, location of insertion and direction of the target gene inserted into the recombinant were correct. After the transfection, over-expression of ZIP10 was found in human breast cancer MCF-7 and MDA-MB-231 cells, while expression of ZIP1 was significantly reduced in the transfected cells. Conclusion     The eukaryotic expression plasmid pEGFP-N1-ZIP10 has been successfully constructed and it can be expressed transiently in MCF-7 and  MDA-MB-231 cells.  The decreased expression of ZIP1 in the transfected cells may indicate the role of ZIP10 in breast carcinogenesis and development.

    Application of sensitized sheep erythrocytes in cell tropism of EV71
    LI Zhi-hui, LI Peng, SONG Nan-nan, YUE Ying-ying, JI Xuan, ZHAO Yuan-hao, MENG Hong
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  33. 
    Abstract ( 1205 )   PDF (1345KB) ( 238 )   Save
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    Objective     To research the cell tropism of enterovirus 71(EV71) using sensitized sheep erythrocytes and explore expression of the EV71 receptor on the surface of passage culture cells.  Methods     Sheep erythrocytes sensitized with EV71 reacted with passage culture cells, and the ability of the cultured cells treated with different methods to absorb the sensitized sheep erythrocyte was observed. Then the cell tropism of EV71 was analyzed and expression of the EV71 receptor on the surface of the cultured cells was explored. Results     Human laryngeal epithelial (Hep-2) cells and rhesus monkey embryo kidney(MA104) cells absorbed the sensitized sheep erythrocytes and the trypsin destroyed the absorption ability of MA104 but not of Hep-2. The absorption of sensitized sheep erythrocytes was not observed in Madin-Darby canine kidney (MDCK) cells. All results agreed well with the results of the EV71 direct infection. Conclusion     This method  proved to be direct and effective to research the cell tropism of virus and expression of the virus receptor on the surface of the cells, and it also has strong feasibility and application value in the research of virology, especially of virus receptors.

    Construction of the mouse T cell immunoglobulin mucin 3(Tim-3) eukaryotic expression vector
    MENG Jing1,LIU Xiao-li2,GUO Chun3,MA Chun-hong3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  37-40. 
    Abstract ( 1195 )   PDF (560KB) ( 453 )   Save
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    Objective    To construct an eukaryotic expression vector of murine T cell immunoglobulin mucin 3(Tim-3), and express Tim-3 in the melanoma cell line B16. Methods   The murine Tim-3 coding region was amplified by reverse transcription PCR (RT-PCR), using RNA from spleen cells as a template. The Tim-3 cDNA was cloned into the eukaryotic expression vector pTARGET by T-A cloning. The recombinant vector was transfected into the melanoma cell line B16 by Lipofectamine. Tim-3 expression was analyzed by RT-PCR and Western blot. Results    The murine Tim-3 expression vector was identified by enzyme digestion and sequencing. RT-PCR and Western blot results showed specific Tim-3 expression in B16 cells transfected with the recombinant vector. Conclusion    The Tim-3 coding region was successfully cloned into the eukaryotic expression vector and highly expressed in the melanoma cell line B16.

    Cloning of the human thioredoxin-1 gene and construction and  identification of its  recombinant vector
    SHI Yan-yan1, DING Shi-gang1, JIANG Su-zhen2, LU Feng-min2, ZHANG Jing1, LIU Lin-na1, WANG Ye1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  41-45. 
    Abstract ( 1042 )   PDF (1309KB) ( 365 )   Save
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    Objective     To clone the human thioredoxin-1 gene (hTrx1) and construct the recombinant vector containing the target gene.  Methods   The cDNA gene of  human thioredoxin was amplified by RT-PCR from gastric cancer cells BCG823 and MNK45, using  specific primers containing double endonuclease digesting sites. The hTrx1 cDNA was then inserted into the EZ-T vector to construct the EZ-T-hTrx1 recombinant vector. The next step was to double digest the EZ-T-hTrx1 recombinant vector and insert the target gene into the pcDNA3.1myc-His to construct the pcDNA3.1myc-His-hTrx1 recombinant vector, which was  identified by double digestion and DNA sequencing.  Results    By double digestion, the hTrx1 cDNA was successfully inserted into the pcDNA3.1myc-His vector; and through DNA sequencing, the sequence of the target gene was inserted and was in accordance with GenBank(NM003329). The pcDNA3.1myc-His-hTrx1 recombinant vector was successfully gained.  Conclusion    The successful cloning of the hTrx1 gene and construction of its recombinant vector pcDNA3.1myc-His-hTrx1 lays the foundation for the study of the biological activity of hTrx1 and mechanism of its function in tumor genesis.

    Effect of the Buchang naoxintong capsule on apoptosis of neurons and inflammatory reaction of microglias after acute cerebral  ischemia-reperfusion injury in rats
    ZHAO Qing1,2, YANG Li-ling1, DU Yi-feng1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  46-49. 
    Abstract ( 1234 )   PDF (1256KB) ( 332 )   Save
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    Objective    To study the effect of the Buchang naoxintong capsule on focal cerebral ischemia-reperfusion injury in rats and to investigate its corresponding mechanisms. Methods    A rat model of middle cerebral artery occlusion (MCAO) and reperfusion was established by I/R .Thirty rats were randomly divided into the sham operation group, the control group and the naoxintong-treated group, with 10 rats in each group. The neurological impairment score was used to evaluate the degree of neurological impairment of rats. Immunohistochemical stainning and Western blot were respectively used to detect expressions of CD68 and IL-1β. TUNEL and Nissl staining were used to detect neuron apoptosis in rats. Results    Activation of microglial cells, TUNEL- and nissl-positive cells, and expressions of CD68 and IL-1β in the naoxintong-treated group were obviously lower than those in the control group. The differences all had statistical significances(P<0.05). Conclusion     The Naoxintong capsule can significantly relieve the drgree of injury of brain tissues and depress apoptosis of neurocytes. The mechanisms could be that the Naoxintong capsule depresses activation of gitter cells which results in release of  inflammation mediators, and then prevents apoptosis of ischemic neurocytes.

    Dynamic changes of nNOS, iNOS and activated Caspase-3 protein  expressions  in the hippocampus of rats with lithium-pilocarpine-induced epilepsy
    CHEN Li-guang1, LIU Bin2, FAN Hai-tao2, JIA Jian-bo2, PANG Qi2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  50-53. 
    Abstract ( 1077 )   PDF (936KB) ( 531 )   Save
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    Objective    To explore the correlation and dynamic changes of protein expressions of nNOS, iNOS and activated Caspase-3 after status epilepticus (SE). Methods       The animal model was established by lithium-pilocarine induction in rats. 2h, 6h, 3d and 7d after SE, variations of expressions of nNOS, iNOS and activated Caspase-3 proteins were determined by immunohistochemistry and immunoblot, respectively. Results    Expression of the nNOS protein significantly increased 2h  after SE(P<0.01), reached its peak at 6h (P<0.01), and decreased thereafter. However, expressions of the nNOS protein on 3d and 7d were still significantly higher compared with the control group (P<0.01). Expression of the iNOS protein was significantly up-regulated 6h after SE(P<0.01), and reached its peak on 3d (P<0.01). Expression of the activated Caspase-3 protein significantly increased 3d  after SE(P<0.01), and reached its peak on 7d (P<0.01). Conclusion     Expressions of nNOS, iNOS and activated Caspase-3 proteins are up-regulated at different levels in the rat hippocampus at different times after SE, suggesting that the activation of Caspase-3 may be involved in the excitotoxic effect induced by nitric oxide synthase (NOS)  and apoptosis could be closely related to neuron injury.

    Relation between CD3+CD16+CD56+ NKT cells and immune  status in patients after renal transplantation
    XIAN Wan-hua1, TIAN Jun1, GUAN Guang-ju2, BI Zheng3, CUI Xian-quan1, SUN Huai-bin1,LI En-gang1, PENG Zhi-guo1, GENG Li-na1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  54-57. 
    Abstract ( 2817 )   PDF (375KB) ( 427 )   Save
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    Objective    To explore the relation of expression of peripheral blood CD3+CD16+CD56+ natural killer T(NKT) cells with immune status and different concentrations of calcineurin inhibitor (CNI) in patients after renal transplantation. Methods    92 patients after renal  transplantation were divided into three groups according to rejection and  concentrations of CNI:patients without rejection in normal concentration(group A,58 cases), patients without rejection in lower concentration (group B,8 cases), and patients with rejection (group C,26 cases). 10 healthy individuals were selected as the controls. CD3+,CD3+CD4+,CD3+CD8+,CD3-CD16+CD56+NK and CD3+CD16+CD56+ NKT cells and human leucocyte antigen(HLA) antibodies were analyzed by flow cytometry.  Results    Percentages of CD3+CD16+CD56+ NKT cells in groups A,B,C and D were (4.29±2.57)%,(4.31±2.08)%,(1.23±1.06)% and (3.98±2.26)%,respectively. Percentages of NKT and NK cells,CD4+/CD8+,and the HLA antibodypositive rate in groups A and B had no significant difference compared with those in group D. The percentage of CD3+CD16+CD56+NKT  cells was lower in group C compared with that in group D (P<0.05). Percentages of NK cells and CD4+/CD8+, and the HLA antibodypositive rate in group C were significantly higher than in group D(P<0.05).   Conclusion     CD3+CD16+CD56+ NKT cells are lowly expressed when rejection occurs,and HLA antibodies occur in these patients. Expression of NKT cells is normal and stable in patients without rejection, and is not affected by CNI concentrations. NKT cells play an important role in immunosuppression in patients after renal transplantation.

    Low molecular weight heparin attenuating non-alcoholic steatohepatitiscaused by high-fat diet in the rat
    SUN Lin-lin, SHI Jun, HAO Jing-hua, REN Wan-hua
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  58-62. 
    Abstract ( 1129 )   PDF (1407KB) ( 278 )   Save
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    Objective     To investigated interference of low molecular weight heparin(LMWH) on non-alcoholic steatohepatitis caused by high-fat diet in the rat and study its mechanism. Methods     Male Sprague-Dawley rats(n=40) were randomly divided into the control group(n=10)and the model group(n=10): the model group was given a highfat diet for 8 weeks;  the low dosage treatment group(n=10) and high dosage treatment group(n=10): one received low doses of LMWH [50IU/(kg·d)] and the other received 200IU/(kg·d) for 2 weeks, after 8 weeks of high-fat diet.   Results    Compared with the model group, a high dose of LMWH[200IU/(kg.d)] significantly inhibited chronic inflammation and attenuated lipid deposits in the liver, while a low dose of LMWH was not significant.  Conclusion     LMWH has an excellent therapeutic effect on non-alcoholic fatty liver disease (NAFLD) by inhibiting chronic inflammation and attenuating lipid deposits in the liver, which is closely related to the dosage. Its mechanism may be that LMWH regulates the level of blood fat and decreases the level of TNFα in the serum.

    Inhibitory effect of short interfering RNA-mediated gene silencing on expression  of VEGF and cell proliferation in murine colon carcinoma cells 
    MA Lan-qing1,GUO Yong-zhang2,ZHANG Hong-bing2,ZHU Zhu2,CHEN Ming-qing3, DUAN Li-ping1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  63-69. 
    Abstract ( 1073 )   PDF (1012KB) ( 345 )   Save
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    Objective    To design three siRNAs targeting against murine VEGF by RNA interference (RNAi) technique and then insert them into the pSliencer4.1 CMV neoexpression vector to investigate the influence on the VEGF gene silencing and biological activity of murine colon carcinoma CT-26 cells, and to explore a novel gene therapy for colon cancer. Methods     ① Three sequences targeting against VEGF were inserted into the pSliencer4.1 CMV neo expression vector after being screened, designed and synthesized. The sequences were identified by restriction enzyme digestion and DNA sequencing.  ② Murine colon adenocarcinoma CT26 cells were divided into 6 groups: the recombinant plasmid V1, V2 and V3 groups(groups V1, V2 and V3), the Lipofectamine group(group Lp), the cell group(group c)and the scrambled group(group Nc). The recombinant vector was transferred into colon carcinoma CT-26 cells using Lipofectamine. Efficiencies of transcription and translation of VEGF were determined by semi-quantitative RT-PCR, Western blot and flow cytometry; and the cell proliferation, cell cycle distribution and cell apoptosis were measured by MTT and flow cytometry.  Results     ① The pSliencer4.1 CMV neo expression vector was successfully constructed and the sequence of the constructed recombinant plasmid was correctly identified by restriction enzyme digestion and DNA sequencing.   ②Expression of VEGF decreased markedly compared with the control groups(P<0.01), and more markedly in groups V1 and V2(P<0.05, groups V1 and V2 vs group V3). ③  The growth of CT26 cells was significantly inhibited in groups V1,V2 and V3 compared with the control groups (P<0.01), and more significantly in groups V1 and V2 (P<0.05, groups V1 and V2 vs group V3). ④ Analysis of the cell cycle distribution showed that the ratio of cells at the G0/G1 phase increased and at S and G2/M phases decreased in groups V1 and V2 compared with group V3 and the control groups (P<0.05). Conclusion     ① The pSliencer4.1 CMV neo expression vector targeting against VEGF was successfully constructed. ②  The pSilencer4.1 CMV neoV1/ V2/ V3siRNA transfecting target CT-26 cells using Lipofectamine has a remarkable gene-silencing effect.

    Effects of sodium butyrate on cell proliferation and expression of ICAM-1 in the HT-29 colon cancer cell line
    LI Wan-quan1, ZHU Xiao-lin2, YU Cheng-gong2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  70. 
    Abstract ( 940 )   PDF (718KB) ( 359 )   Save
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    Objective    To investigate the effects of sodium butyrate on HT-29 colon cancer cell proliferation and expression of intercellular adhesion molecule 1 (ICAM-1) on the cell surface, and to explore the mechanism by which sodium butyrate affects growth of HT-29 cells. Methods    Effect of sodium butyrate on the proliferation of HT-29 cells was detected by MTT[1-(4,5-Dimethylthiazol-2yl)-3,5-diphenylformazan] assay. The cell cycle, cell apoptosis and expression of ICAM-1 induced by sodium butyrate were measured by flow cytometry. Results    In a time-and dose-dependent manner, sodium butyrate significantly inhibited the growth of HT29 cells, increased the proportion of G0/G1 cells and the proliferation index, decreased the proportion of S phase cells, and induced cell apoptosis. Also, sodium butyrate down-regulated expression of ICAM-1. Conclusion    Sodium butyrate can inhibit the proliferation of HT-29 cells. The mechanism may relate to arresting the cell cycle, inducing cell apoptosis and down-regulating expression of ICAM-1 on HT-29 cells.

    Research of nuclear factor-κB siRNA to prevent proliferation  and restenosis of vein grafts
    MENG Xiang-bin1, BI Yan-wen1, FENG Jin-bo2, ZHANG Jian-ping3, YUE Wei-ming4, SONG Guang-min1, SUN Wen-yu1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  74-80. 
    Abstract ( 952 )   PDF (1499KB) ( 267 )   Save
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    Objective    To investigate the effects of nuclear factor κB small interfering RNA (NF-κB siRNA) on intimal proliferation and expressions of inflammatory factors in autologous vein grafts. Methods    A total of 80 male Wistar rats(weight, 300g to 350g)were randomly divided into four groups: 1) Group A( n=10), normal group without transplantation; 2)Group B(n=18), vein transplantation only; 3) Group C (n=26),  vein transplantation but the graft was processed by blank plasmid liposome complexes; 4) Group D(n=26), vein transplantation but the graft was transfected by NF-κB siRNA liposome complexes. The transplantation was to replace an  abdominal aortic vein with the autologous right external jugular vein.  At four time points (3rd ,7th,14th,21st day),  autografts were processed to determine intima hyperplasia, PCNA, MCP-1 mRNA, TNF-α mRNA and NF-κB p65 protein. Results    Intimal hyperplasia and VSMC obviously proliferated in groups B,C and D. At the 3rd day, expressions of MCP-1mRNA and TNF-αmRNA in group D was significantly lower than group C (P<0.05). At the 7th day, the levels of NF-κB p65 protein in group B and C were significantly higher than in group A (P<0.05). Compared with group C, the levels of NFκB p65 protein in group D were lower(P<0.05). Conclusion     Expressions of MCP-1 mRNA and TNF-α mRNA have some relationships with NF-κB p65 protein. The proliferation of vascular endothelial cells and expression of inflammatory cytokines are changing and correlative in the vein graft. Transfection of NF-ΚB siRNA liposome complexes on the autologous vein graft can inhibit its proliferation and reduce expressions of NF-κB, MCP-1 and TNF-α.

    Effects of ZnCl2 and Zn-EDTA on formation of the epidermis granular layer in the mouse tail
    LIU Zhen-jun1,CAO Jun1,FU Jun2,CUI Xi3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  81-83. 
    Abstract ( 1115 )   PDF (470KB) ( 278 )   Save
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    Objective    To study effects of ZnCl2 and Zn-EDTA on formation of the epidermis granular layer in the mouse tail.  Methods    The models of tail scale epidermis were established by using healthy male Kunming mice. The mice were randomly divided into the  negative and positive control groups, low, middle and high dosage ZnCl2 groups, or and low, middle and high dosage Zn-EDTA groups. According to the protocol, normal saline, methotrexate, ZnCl2 or Zn-EDTA were intragastrically given to each group for 12 consecutive days. 1d after the last administration, mice were sacrificed and pathological sections of the tail scale epidermis were made to count the number of scales with the granular layer in 100 scales under a light microscope. Results     Different doses of ZnCl2 and ZnEDTA all significantly promoted the formation of the granular layer in the mouse tail scale epidermis (P<0.05), while no significant difference was found compared with the methotrexate group, which acted as the positive control(P>0.05). Conclusion    ZnCl2 and Zn-EDTA promote skin keratinization of the mouse tail scale.

    Neurodegeneration in different brain regions of neonatal  rats damaged by hypoglycemia 
    ZHOU Dong1, QIAN Jing2, CHANG Hong3, LI Fu-hai1, SUN Ruo-peng1
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  84. 
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    Objective   To establish a reliable animal model of brain injury induced by neonatal hypoglycemia and observe the  corresponding neurodegeneration in different brain regions of neonatal rats. Methods    Newborn Wistar rats were randomly divided into: insulin-treated rats with short hypoglycemia (INS-S group, n=40), insulintreated rats with prolonged hypoglycemia (INS-P group, n=40), fasting rats (FAS group, n=40) or control rats (CON group, n=40). Neonatal hypoglycemia was produced by fasting or insulin injections (15U/kg, hypodermic injection) on postnatal day 2 (P2), 4 (P4), and 6 (P6). Double labeled immunofluorescent with neuronal nuclear protein (NeuN) and FJB was performed to examine the neuronal degeneration in the rat brain 2h, 6h, 1d, 3d, 7d and 14d after the third hypoglycemic insult. Results   No FJB+ cells were detected in INS-S, FAS and CON rats at any designated time point, but numerous FJB+ cells were detected in INSP rats at 6h, 1d, 3d and 7d after the third hypoglycemic insult. Double labeled immunofluorescent with NeuN showed these FJB+ cells all presented NeuN immunoreactivity. There were numerous FJB+ cells in the parasagittal cortex, piriform cortex, hypothalamus, thalamus and dentate gyrus. Conclusion    Insulin hypodermic injections in newborn rat can establish a reliable animal model of brain injury damaged by neonatal hypoglycemia. Repetitive and profound neonatal hypoglycemia can result in extensive neurodegeneration, in which the neurons of the cortex, thalamus, hypothalamus and dentate gyrus are more vulnerable.

    Association of urinary albumin-to-creatinine ratio with femoral arterial stiffness in essential hypertension
    MA Wei-hong1,2, BU Pei-li2, WANG Xiu-ling2, LIU Rong2, LIU Jun-ni2, LI Chang-jun2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  90. 
    Abstract ( 1094 )   PDF (551KB) ( 662 )   Save
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    Objective    To explore the association of microalbuminuria(MA) with femoral arterial stiffness and sensitive indicators of early target organ damage in hypertensive patients. Methods     128 individuals were divided into three groups: the healthy controls (n=35),patients with normal microalbuminuria (group A, n=50), and microalbuminuria (group B,n=43). Body mass index (BMI), fasting blood glucose (FPG), triglycerides (TG), total cholesterol (TC), serum creatinine(Cr), glomerular filtration rate (GFR), high density lipoprotein cholesterol (HDL),low density lipoprotein cholesterol(LDL),blood urea nitrogen(BUN),systolic pressure(SBP),diastolic pressure(DBP) and pulse pressure(PP) were measured in all subjects. Urinary albumin-to-creatinine ratios(ACR)were measured with immunoturbidimetry; femoral arterial intima-media thickness (IMT), peak systolic velocity (PSV), end diastolic velocity (EDV), systolic diameter (Ds) and diastolic diameter (Dd) were measured with ultrasonography; stiffness, tensityand distensibility of the femoral artery were calculated; pulse wave velocity(cfPWV) were measured with an arterial elasticity meter.  Results    Compared with the controls, ACR, cf-PWV and stiffness in group A and group B were significantly increased (P<0.05); tensity  and distensibility in group B were significantly decreased (P<0.01); ACR was positively related with PWV and stiffness (P<0.01) and negatively related with femoral tensity and distensibility (P<0.05).  Conclusion     ACR, cf-PWV and femoral arterial stiffness are non-invasive and sensitive indicators,which could predict early target organ damage and cardiovascular risk in hypertension.

    RhD isoimmunization analysis of 304 RhD-negative pregnant and lyingin women  
    GUO Wei1, XU Qun2, SHAO Chao-peng3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  94-97. 
    Abstract ( 1412 )   PDF (343KB) ( 387 )   Save
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    Objective     To analyze the RhD isoimmunization state of 304 RhD-negative pregnant and lying-in women, explore factors which affect production of the antiD antibody in them, and set up a correct prenatal testing method for RhD haemolytic disease of the fetus and newborn. Methods   ABO and RhD blood types of pregnant and lyingin women and their husbands were identified by the standard serological method. For RhD-negative women, the D antigen was further tested with the indirect antiglobulin test to exclude or confirm weak D or partial D types. The RhC, c, E and e antigens were typed in all RhD-negative women and their husbands. The irregular antibody was screened in all RhD-negative women, and if the result was positive, the specificity and titer of the antibody were determined. At the same time, if phenotypes of the RhD-negative women with the anti-D antibody were Del type or not was determined with the PCR-SSP method.  Results     Among 3,975 pregnant and lying-in women, 304 were confirmed to be RhD-negative, of whom 29  produced the anti-D antibody; 24 carried compatible ABO antigens with their husbands(82.76%); 5 carried incompatible ABO antigens with their husbands(17.24%) . The production rate of the RhD antibody in D-negative pregnant and lyingin women was 9.54% (29/304) . All 29 RhD-negative women with anti-D were excluded from carrying the Del phenotype with the molecular biological method. Conclusions     RhD isoimmunization is affected by many factors. Pregnant women with the Del type seldom produce the anti-D antibody. The anti-D level should be timely and regularly tested among pregnant women. For women who have produced the anti-D antibody,  the antibody level should be closely detected during pregnancy, which will  provide evidence for clinical treatment.

    Application of axial laminar screws combined with atlantal lateralmass screws fixing   method in the surgery of complicated atlantoaxial dislocation
    ZHANG Yuan-kai, LIU Pei-lai, LI De-qiang, LI Ming
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  98. 
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    Objective     To explore the application of axial laminar screws combined with the atlantal  lateral mass screws fixing method in surgery of complicated atlantoaxial dislocation. Methods     6 cases of atlantoaxial dislocation complicated by dysplasia or comminuted fracture of the axial pedicle root were involved in the study. Axial laminar screws combined with the atlantal lateral mass screws fixing method was performed on the patients, followed by fusing of the atlantoaxial joint using bone grafts. Results    Satisfying reduction and stability of the cervical spine were shown in the X-ray film and CT image instantly after surgery. Adequate atlantoaxial fusion was shown in the X-ray film during follow-up. No evidence indicated  vertebral artery or spinal cord injuries. Conclusion     Axial laminar screws combined with the atlantal lateral mass screws fixing method has a perfect effect on the surgery of atlantoaxial dislocation complicated by dysplasia or comminuted fracture of the axial pedicle root.

    A  case-control study on influencing factors of esophageal squamous cellcancer in males of Feicheng City
    ZHANG Hao1,ZHOU Ying-Zhi2, DIAO YuTao3, LI Hui-Qing3, ZHOU Rui-xue4, ZHAO De-li4, LEI Fu-hua4
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  102-105. 
    Abstract ( 1136 )   PDF (343KB) ( 286 )   Save
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    Objective    To investigate the influencing factors of esophageal squamous cell cancer(ESCC) in males of Feicheng City, a high ESCC incidence area of Shandong Province, China. Methods    In a high incidence community of ESCC, a cohort of residents aged from 40 to 69 were examined for esophageal lesions. The case group included 64 male cases of ESCC diagnosed by staining endoscope and pathogenic examination and 116 male inpatients of Feicheng People′s Hospital,and 220 males with normal esophageal mucosa determined by gastric endoscope served as the control group. The associations of family history of esophageal cancer, smoking and drinking with esophageal cancer were analyzed, and OR and 95%CI were used as the association indicators. Results    Age and education level were regarded as adjusting factors. No significant association was found between family history of esophageal cancer and ESCC. When smoking index=0 was used as the base line(OR=1.0), the OR was 1.552 (95%CI:0.844-2.851) and 2.273 (95%CI: 1.2484.142)  for smoking index <550 and ≥550 respectively; when drinking index=0 was used as the base line(OR=1.0), the OR was 2.887  (95%CI: 1.4685.679) and 3.281 (95%CI: 1.7806.045) for drinking index <130 and ≥130 respectively; when no-smoking and no-drinking was used as the base line(OR=1.0), the OR was 2.907 (95%CI: 1.3726.162) for both smoking and drinking.  Conclusion    Smoking and drinking are risk factors of ESCC for males with dose-effect relationships, and there is synthetic effect between smoking and drinking on esophageal cancer. Abstinence from smoking and drinking has important preventive effect against esophageal cancer in high incidence areas of ESCC.

    Clinical application of three-dimensional transrectal  ultrasonography to  the diagnosis of perianal infections
    Lv Yan-feng, WANG Jian-xin, LI Tao, DING Ke, BEI Shao-sheng, YU Hua-long
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  106. 
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    Objective    To evaluate the role of three-dimension transrectal ultrasonography in locating the internal opening,  trend of fistulas, pus-filled cavity and occult focus in diagnosing perianal infections.  Methods   127 cases of anal fistula and 48 cases of perianal abscess were examined by three-dimensional endoluminal ultrasonography.  According to the characters of the sonogram and echogram, it located the internal opening, traced the trend of fistulas, and detected the pusfilled cavity′s location and the occult focus. Results    In 151 cases the internal opening was located with an accuracy rate of 86.3%(151/175); in 90 cases one fistula was located, and in 37 cases two fistulas were located. 55 cases of abscess were examined, including 9 cases of sub-cutaneous abscess, 3 cases of sub-mucosal abscess, 4 cases of mucosa subcutaneous abscess, 16 cases of low intramuscular abscess, 11 cases of high intramuscular abscess, 3 cases of ischial rectal space abscess, and 2 cases of pelvic space abscess. All  the above were confirmed by surgery. Conclusion    Three-dimensional endoluminal ultrasonography in diagnosing perianal infections can accurately locate the internal opening, display the trend of fistulas, and effectually detect the pus-filled cavity′s location and the occult focus. And it can provide essential diagnostic criteria for choosing the clinical therapy method.

    Relationship of serum sodium to complications and  prognosis of liver cirrhosis
    ZHANG Jun-yong1, KUAI Jing-hua1, QIN Cheng-yong1, LI Wei2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  109-112. 
    Abstract ( 950 )   PDF (512KB) ( 292 )   Save
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    Objective     To explore the relationship of serum sodium to complications and prognosis in patients with end stage liver cirrhosis.  Methods       The data from decompensated cirrhosis patients in our hospital were selected to be analyzed. Serum sodium concentration was classified. The relationship between incidences of complications and serum sodium concentration was analyzed. Mortality in different sodium concentrations was determined by the Kaplan-Meier survival analysis. Using the model of end stage liver disease(MELD), characteristics of serum sodium in different MELD values were analyzed. The relationship between serum sodium and prognosis was analyzed by Cox proportional hazard regression. Results     In 467 patients, the incidence of hyponatremia (<135mmol/L) was 50.54%. Except for bleeding from the digestive tract, incidences of other complications, such as hepatorenal syndrome, hepatic encephalopathy and spontaneous peritonitis, were higher in hyponatremia than in a sodium concentration ≥135mmol/L (P<0.05). When the concentration was ≤125mmol/L, the incidence of a large amount of ascites was 88.24%, which was obviously higher than that at other concentrations(P=0.000). Mortality of cirrhosis patients was increased when the serum sodium concentration decreased. The Kaplan-Meier survival analysis showed that mortality was statistically different inconcentrations of ≤125mmol/L, (125135)mmol/L and ≥135mmol/L (P=0.000). Sodium concentration could be one of the independent prognostic predictors by the Cox proportional hazard regression(P=0.000).  Conclusion    The complications closely correlate with hyponatremia, which can be one of the prognostic predictors in patients with end stage liver disease.

    Inhalation of long-term glucocorticoid and long-acting β2-agonists in improving lung function and airway inflammation in chronic obstructive pulmonary disease
    YANG Lin-hong,WANG Jun,CHEN Wen-jing,LI Jia-zhao
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  113. 
    Abstract ( 1300 )   PDF (379KB) ( 282 )   Save
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    Objective    To explore the improvement of lung function and airway inflammation by using glucocorticoid and long-acting β2-agonists (CABA)or by using a single drug for chronic obstructive pulmonary disease(COPD). Methods    60 moderate COPD patients were randomly divided  into four groups: the treatment group 1 inhaled shamoteluo / fluticasone and formoteroln, the treatment group 2 inhaled formoterol, the treatment group 3 inhaled budesonide, and the control group used ammonia Tualkali and short-acting β2 receptor agonists on-demand applications. Blood gas analysis, FEV1.0%, serum tumor necrosis factor (TNF-α) and transforming growth factor (TGF-β) were determined before the treatment and at 3, 6, 12, 24 months during the treatment. Results    Acute attacks in the treatment groups were significantly reduced. Compared with the control group there were significant differences (P<0.05), and in the treatment group 1, the difference was the most significant (P<0.01). At 6 months, FEV1.0%, TGF-β, and TNF-α had more significant differences in the treatment groups than in the control group (P<0.05), but there were no significant difference between the  24 months  group and the 12 months(P> 0.05), while there were no significant changes both before and after the treatment in the treatment groups (P> 0.05). PaO2 of the four groups had no significant changes from that before the treatment (P>0.05). Conclusion     Inhalation of both longterm glucocorticoid and LABA, or single glucocorticoids or single LABA can reduce acute attacks of COPD, improves patients‘ lung functions, and reduces concentrations of serum TGF-β and TNF-α. The effect of inhaled glucocorticoid in combination with LABA is more valuable than single inhalation of glucocorticoids or LABA.

    Splenectomy for idiopathic thrombocytopenic purpura: prognosis analysis of 75 cases
    FENG Rui1, ZHANG Hai-xia2,LI Hong-mei3
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  117-120. 
    Abstract ( 1061 )   PDF (334KB) ( 283 )   Save
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    Objective      To investigate the efficacy of splenectomy on patients with idiopathic thrombocytopenic purpura(ITP), and the influence of the preoperative clinical indexes on the efficacy.Methods     Clinical data from 75 (11 men, 64 women) ITP patients with splenectomy were reviewed, and grouped according to the sex, age, course of disease, platelet count, megalokaryocyte number, spleen size, and treatment outcome of glucocorticoid. The relations of each factor with splenectomy efficacy were compared and analyzed.  Results     Among 75 cases, 42 with excellent response (56.0%), 18 with good response (24.0%), 9 with progression(12.0%),6 with no response(8%). 8 patients relapsed after surgery. The response rate (the percentage of cases with excellent and good response in all cases) was 80.0%. The response rates were 81.8% and 79.7% in male and female (P>0.05), 80.3% and 77.8% in patients with and without spleen enlargement (P>0.05), 85.4% and 70.4% in patients younger and older than 40 years(P<0.05), 85.2 % and 66.7% in patients responsive and irresponsive to glucocorticoid therapy (P<0.05), 85.7% and 63.2% in patients with the bone marrow megakaryocyte number ≥ and < 7-35/(3×1.5)cm2 (P<0.05), 86.0%and 68.0% in patients with preoperative platelet count > and <20×109/L (P<0.05), and 84.5% and 64.7% in the disease course below and over 6 months, respectively (P<0.05). Conclusion     There is no relation of splenectomy efficacy with the gender or spleen size. However, the age, course of disease, bone marrow megakaryocyte number, platelet count, and the response to glucocorticoid are obviously relative to the efficacy of splenectomy.

    Short-term curative efficacy of cytokine-induced killer cells on patients  with gynecological malignant carcinoma
    ZHANG Bei1,2, YAN Li2, ZHOU Ting2, RONG Feng-nian2
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  121. 
    Abstract ( 1070 )   PDF (460KB) ( 377 )   Save
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    Objective     To observe the short-term curative efficacy of transfusing autogeneic cytokine-killer cells(CIK) cultured in vitro on patients with gynecological malignant carcinoma. Methods     Blood samples isolated from 8 patients with gynecological malignant carcinoma were induced in vitro to obtain CIK, which were then transfused into the patients. The short-term curative efficacy was evaluated by comparison of the immunological function, level of CA125 in serum, Karnofsky performance score(KPS) and change of clinical symptoms before and two months after transfusion. Results      After transfusion, both the ratio of CD4+/CD8+ and percentage of CD3-CD16+CD56+ in peripheral blood were increased(P<0.05),while the percentage of CD4+CD25+ was decreased(P<0.05). The level of CA125 in serum was decreased in 5 cases. The KPS was increased in one case. Clinical symptoms were ameliorated in 5 cases. At the same time, no adverse reaction was observed. Conclusion     Transfusing autogeneic CIK has some short-term curative efficacy on gynecological malignant carcinoma.

    Contamination of ochratoxin A in millet and maize sold in Jinan
    YANG Yan-you1, GAO Wen-hua2, WEN Hong-ling3, PU Shuang-shuang3, SONG Yan-yan3, XU Hong-zhi3, ZHAO Li3, LI Feng-qin4
    JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(11):  125-128. 
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    Objective     To determine the contamination of ochratoxin A in millet and maize commercially available in Jinan.  Methods     An optimization study of the extraction solvent, testing conditions and recovery rates in detection of ochratoxin A in millet and maize by ELISA was carried out. Millet and maize samples collected from 5 districts of Jinan were analyzed under optimum conditions. Results      Optimal ochratoxin A extraction was obtained when the extracting agent contained 5% sodium chloride and 60% methanol. For all the samples, the ochratoxin A detectable rate in millet was 41%, while in maize it was 38%. The ochratoxin A detectable rate in millet from supermarkets was 56%, which was significantly higher than that in the millet samples from markets (26%) (P<0.05). The ochratoxin A concentration level ranged between 0.40 and 4.45μg/kg, and the median was 1.67μg/kg. No significant difference in the ochratoxin A pollution level between millet and maize was found (P>0.05). Ochratoxin A contents in all samples were below the tolerance limit set by the state for ochratoxin A in grain. Conclusions     The millet and maize commercially available in Jinan are contaminated by ochratoxin A to some extent, but the ochratoxin A content in the detected samples is below the tolerance limit.