Table of Content

  

24 April 2006
Volume 44 Issue 4

Articles


Articles

Effect of dietary fat intake on human serum apolipoprotein AIV levels

XU Guifa,LI Hui,LIU Gensheng,SHI Mai,LI Hui

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(4):  325-327. 

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Objective: To investigate the effect of dietary fat intake on human serum apolipoprotein(apo) AIV levels and further illustrate the regulation of food intake and bodyweight gain. Methods: This investigation was conducted in 389 subjects aged 770 years from Zibo and Jinan city in Shandong Province. The serum apoAIV concentrations were measured by enzymelinked immunosorbant assays (ELISA). Dietary intake was estimated from 3day food record. Statistical analyses were performed with SPSS 12.0. Results: Serum apoAIV levels were negatively correlated with dietary fat, fiber, carbohydrate, and protein intake（P<0.05）. Humans chronically consuming a highfat diet initially developed significantly lower serum apoAIV levels than those of a lowfat diet. Conclusion: A highfat diet intake is the primary factor to affect serum apoAIV levels.

Prevalance of human herpesvirus 8（HHV8）IgG and its associated risk factors in blood donors from Jinan region

QI Mei,ZHAO Weiming,ZHOU Yabin,LUAN Yi,BIAN Jifeng,WANG Hong,

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(4):  328-331. 

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Objective: To estimate the seroprevalence of HHV8 infection and its correlation with hepatitis B virus (HBV), hepatitis C virus (HCV) and syphilis infection among blood donors in Shandong region. Method: Five hundred and twenty serum were tested for HHV8 IgG antibody with ELISA by using HHV8 ORF K8.1 peptides as antigen. Results: The mortality of positive HHV8 IgG antibody in blood donors from Jinan region was 5.962%（31/520）, and had a trend of increase with the blood donors′ increasing age, but it was not associated with exposure to HBV, HCV or syphilis. Conclusion: There is a certain rate of HHV8 infection in blood donors of Shandong. It is necessary to screen blood donors for HHV8.

Protective effects of NGF and GM1 on primary sensory neurons in SD rat with sciatic nerve injury

HUANG Fei,WANG Huaijing,XING Yi,GAO Wei,LI Yonggang,XING Ziying,LI Zhenzhong

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(4):  332-335. 

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Objective: To observe the protective effects of nerve growth factor (NGF) and monosiaganglioside (GM1) on primary sensory neurons in rats with sciatic nerve injury. Methods: Ninetysix SD rats were randomly divided into normal saline control group(A), NGF group(B), GM1 group(C) and NGF + GM1 group(D). The sciatic nerve was incised with a 5 mmdefect. The two ends of nerve were connected with a 10.0mm length silicagel tube of 2.0 mm bore. Dorsal root ganglia (DRG) was removed from each animal at different time after treated with saline, NGF, GM1 and NGF+GM1 respectively. Results: At the 4th week after operation,the number of neurons and nerve conductive velocity(NCV) increased significantly(P＜0.01) in the treatment groups than in the control group, and it increased significantly(P＜0.01) in group D than that in group B and C. At the 8th week after operation, the number of neurons and NCV increased significantly(P＜0.01) in the treatment groups than that in the control group; whereas there were no significant difference between each treatment groups. Conclusion: Using NGF and GM1 simultaneously may avoid the necrosis of neurons and accelerate the regeneration of DRG neurons when peripheral nerves are damaged, which is superior to that using NGF or GM1 alone.

Construction of expressive vector containing double suicide genes controlled by MDR1 promoter and its expression in K562/A02 cell

WANG Xiangling, JI Chunyan, MA Daoxin, ZHAO Jianqiang, HOU Ming, YU Haiqing, ZANG Shaolei

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(4):  336-340. 

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Objective: To construct expressive vector containing double suicide genes targeted by MDR1 promoter for the purpose of targeted gene therapy for MDR leukemia. Methods: The DNA fragment of MDR1 promoter was amplified from genome DNA of K562/A02 cells with PCR and was inserted into the upstream of CD-TK to construct pcDNA3MDR1PromoterCDTK.This recombinant vector was transfected into K562, K562/A02 cells by means of liposome. PCR and RTPCR were resorted to identify the integration and expression of CD and TK genes. Results: The length and sequence of MDR1 promoter amplified by PCR were confirmed by DNA sequencing. The pcDNA3MDR1PromoterCD-TK expression vectors were constructed successfully. PCR proved double suicide genes were integrated into K562/A02 and K562 cells. RT-PCR revealed that CD and TK genes expressed in K562/A02/CD-TK cells, whereas not in K562/CDTK cells. Conclusion: Construction of expressive vector containing double suicide genes targeted by MDR1 promoter and its specific expression in K562/A02 cell provide a sound basis for targeted gene therapy for MDR leukemia.

Effects of losartan on the platelet activation and expression of angiotensin Ⅱ AT1 receptor mRNA after vascular balloon injury in rats

WU Yi-nan,GE Zhi-ming,LI Yong-hong,LI Fang,ZHANG Cheng-qiu,ZHANG Yun

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(4):  341-344. 

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Objective: To investigate the process of neointimal formation, level of platelet activation, expression of angiotensinⅡ AT1 receptor mRNA and effects of losartan on them after vascular balloon injury in rats. Methods: Fortyeight male Wister rats were randomly given aortic balloon injury by 2F balloon catheters (n=24), aortic balloon injury plus losartan (30?mg•kg-1•d-1) from 1 day before injury to 28 days after injury, and compared with the controls (n=24). The process of intimal thickening, number of platelet GMP140 and level of AT1 receptor mRNA were investigated at day 3,7,14 and 28 after injury by histological method, radioimmunological method and RTPCR technique, respectively. Results: ① The expression of AT1 receptor mRNA increased significantly at day 3 after balloon injury and remained high at day 14 (P<0.01). ② The number of platelet GMP140 was high at day 3 and began to decrease at day 7 after injury（P<0.01）. ③ The migration and proliferation of vascular smooth muscle cells(VSMC) had existed at day 3 after balloon injury.The thickening of intima happened at day 7 after injury　(P<0.01) and acted more significant at day 14　(P<0.01). The proliferation of VSMC decreased at day 28, but extracellular matrix increased and intimal thickening continued　(P<0.01).④ In losartan treated group，the number of platelet GMP140，proliferation of VSMC and intimal thickening reduced　(P<0.01), but the level of AT1R mRNA increased　(P<0.01). Conclusion: The expression of AT1R mRNA and the number of platelet GMP140 increase in the process of intimal thickening after balloon injury. Losartan inhibits the activation of platelet and intimal thickening，but upregulates the expression of AT1R mRNA.

Construction of RNA interfering expression vector directed against Nrf2

YANG Xiaoyun,LI Yanqing,LIANG Xiaohong,GUO Yuting,YUAN Junhua,ZHANG Yan,ZHU Qiang

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(4):  345-350. 

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Objective: To construct a RNAi expression vector aimed at human Nuclear factor E2 p45related factor 2 (Nrf2) gene and it to study the chemoprevention for colon cancer. Methods：Two sequences targeting the ORF of Nrf2 were cloned into the RNA polymerase III based expression vector pSUPER. These recombinants were transfected into HT29 cells. Fluorescence microscope and flow cytometry were used to determine the lipfectin transfection efficiency after being transfected with pEGFPN1 plasmids. The stable cells were selected in medium 48hours after pEGFPN1cotransfected with G418. The expression of Nrf2 was assayed using RTPCR and Western blotting. RTPCR analysis of UGT1A mRNA was performed on the stable cells. Results：The construction of the recombinant expression vector pSUPERNrf2A1，B1 and its control vector pSUPERNrf2A2，B2 was successfully confirmed by the results of enzyme digestion, electrophoresis and sequencing. The transfection efficiency was 30％～75%. The ability of these vectors inhibiting Nrf2 in a transient and stable expression experiment in HT29 cells was compared.Importantly, pSUPER Nrf2B1 was able to significantly knockdown Nrf2 expression. pSUPERNrf2A1 only had a moderate activity, whereas pSUPER Nrf2A2，B2 were inactive in this assay. Moreover, activities of UGT1A was reduced by 20％～30％ in the stable cells transfected with pSUPERNrf2A1，B1 vector. Conclusion：siRNA expression mediated by the pSUPER vector causes efficient, stable, and specific downregulation of Nrf2 gene expression，suggesting the suppression of Nrf2 gene expression results in downregulation of the constructive expression of UGT1A gene.

Cloning and screening of the recombinant plasmids

ZHU Yuhua,ZHANG Chunqing,REN Wanhua,MA Yanli,ZHAO Youan

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(4):  351-356. 

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Objective：To clone the recombinant plasmids expressing connective tissue growth factor(CTGF) short hairpin RNA(shRNA) by Pgenesil1 plasmid, and to screen the highly efficient shRNA. Methods：Three pairs of two DNA sequences containing small hairpin structure and one pair of unspecific control sequence were designed and synthesized respectively. The complement forms were obtained by annealing and inserted into plasmid Pgenesil1 with U6 promoter, and the recombinant plasmids were transformed into DH5α strain. Finally the plasmids identified by restriction enzyme were used for sequence analysis. Four pairs of recombinant plasmids being transfected into HSCT6,the expression of CTGF mRNA level was determined by reverse transcriptionpolymerase chain reaction 24 hours later. Results: The CTGF shRNAs were successfully inserted into plasmid Pgenesil1. The recombinants were identified by endonuclease digestion and sequencing. Compared with the controls, the expression of CTGF mRNA level in HSCT6 transfected with CTGF shRNA recombinants was markedly downregulated by (74±5)%(P<0.01) and (61±3)%(P<0.05) respectively in two groups of recombinant plasmids, wherease it had not any significant changes in another shRNA and unspecific shRNAtransfected and only with transfection reagent to HSCT6. Conclusion: Three pairs of recombinants expreessing CTGF shRNA are successfully constructed, shRNA sequences potently inhibiting CTGF are screened out successfully too, which lends itself to new gene therapy for liver fibrosis.

Hepatic tissue expression of HLAI in patients with chronic hepatitis B virus and hepatocellular carcinoma

CHENG Baoquan,XIE Yinghui,LIU Chuntao,ZHONG Ning,FAN Wei,ZHANG Shangzhong

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(4):  357-359. 

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HObjective: To study the expression of human leucocyte antigen I（HLAⅠ） in patients with hepatitis B virus infection and hepatocellular carcinoma. Method: HLAI in 51 liver tissues of viral hepatitis B and 43 tumor tissues of hepatocellular carcinoma was examined by flow cytometry. Results: The expression of HLAI was lower in tumor tissue of hepatocellular carcinoma(36.47±19.38) than that in liver tissue of chronic viral hepatitis B(79.54±41.55, P＜0.01)and normal liver tissues(81.86±46.53, P＜0.01), but no significant difference was found between the normal liver tissues and tissues of chronic viral hepatitis(P＞0.05). Moreover, the expression of HLAI in tumor tissues with lymphnode involvement was lower than in tumors without involvement(22.83±11.59 vs 46.28±23.16) (P＜0.05). There was a reverse relation between the expression of HLAI and the level of serum alpha fetoprotein (r=-0.741?1, P＜0.01), but a positive relation was found between with histological differenciation grading(r=0.805?1, P＜0.05). Conclusion: Low expression of HLAI is relation to development and metastasis of hepatocellular carcinoma.

Expression of PDGF and its receptor on pancreas fibrosis in rats

WANG Dongsheng,LIU Qian,KOU Yan

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(4):  360-363. 

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Objective: To study the expression and significance of PDGF and its receptor in the process of pancreas fibrosis. Methods: The animal model of chronic pancreatitis was established by injection of 2% TNBS to rat′s pancreatic duct. Expressions of PDGF, PDGFR and αSMA were determined in pancreas tissues in different stages by using immnnohistochemistry technique. Results: Expressions of PDGFB, PDGFRβ and αSMA were stronger in model than in control group on the fourth week. There was a positive correlation between αSMA and PDGFB and PDGFRβ, and no correlation between αSMA and PDGFA and PDGFRα. The expressions of PDGFB and PDGFRβ were stronger than those of PDGFA and PDGFRα with a significant difference in model group. Conclusion: PDGFB and PDGFRβ are mainly to act play a key role in pancreas fibrosis and development.

Effects of competitive blood flow from differently stenotic coronary artery on internal mammary artery graft flow

FU Qiang,BI Yanwen,YU Jianmin,SUN Wenyu,PANG Xinyan,LIU Wei,QIAO Li

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(4):  364-367. 

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Objective:To study the effects of competitive blood flow from differently stenotic coronary artery on internal mammary artery graft flow. Methods: Coronary artery bypass grafting (CABG) was conducted in 15 Chinese miniswines using left internal mammary artery(LIMA) anastomosed to left anterior descending artery (LAD). The proximal LAD was mad differently stenotic (0%,30%,50%,75%,90% and 100%) through a flow occlusion and the mean flow of LIMA and LAD proximal and distal to the anastomosis were respectively measured through the transit time flow measurement (TTFM). Results:The mean flow of LIMA was (9.75±1.45), (11.63±1.69), (15.63±2.26), (19.75±2.37), (23.50±2.34) and (26.75±2.11)ml/min, respectively. The mean pulsatility index (PI) value was respectively 4.4±1.7, 4.1±1.6, 4.2±1.9, 3.7±1.8, 3.3±1.6 and 2.5±1.4. Themean flow of LIMA when the stenosis of the proximal LAD was 90% or 100% was significant higher than that of LIMA when the stenosis of the proximal LAD was 0%, 30% or 50%（P<0.01）. When the proximal LAD was not fully occluded, there were dual flow in LIMA. There were no difference in LAD flow distal to the anastomosis. Conclusion: The competitive flow from patent LAD does exist and has important effects on IMA graft flow and may be an important cause of IMA graft failure.

Detection of micrometastasis in the bone marrow of patients with esophageal carcinomas and its clinical implications

LV Yingyi, CHEN Jinghan, PENG Zhongmin, MENG Long

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(4):  368-371. 

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Objective：To study the micrometastasis in the bone marrow of patients with esophageal carcinomas and its clinical implication. Methods：CEA mRNA expression in bone marrow of rib was detected in 43 patients with esophageal carcinoma and 17 patients with esophageal benign lesions as controls by RTPCR. Results：The positive CEA mRNA expression was 34.9 %(15/43) in bone marrow of patients with esophageal carcinoma and not found in patients with benign lesions (P＜0.05). No correlation was observed between the expression and sex,age, histological type,cancer cell differentiation, but close correlation between the expression and depth of invasion,lymph node metastasis(P＜0.05）. Conclusion：RTPCR amplification of CEA mRNA is a sensitive and specific method for detection of bone marrow of rib segments in patients with esophageal carcinoma. The detection of CEAmRNA in bone marrow is helpful in comprehensive judging the extent and the prognosis, leading to an early diagnosis and treatment for patients with subclinical metastasis in bone marrow.

The pilot study on realgar for glioma

PANG Qi,WANG Hanbin,GE Mingxu,SI Zhi-chao,ZHU Yu-fang,ZHANG Kang,WEI Guang-fu,ZHANG Zhi-ming

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(4):  376-379. 

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Objective: To investigate the potential effects of realgar on the glioma cells both in vitro and in vivo, and evaluate its role as a novel agent against human glioma. Methods: The 9 L glioma cells were cultured in suitable medium, cell proliferation inhibitory assay was employed to observe the reaction of these cells, including the morphological changes and cytotoxicity by realgar suspension at a certain concentration; local injection or oral administration or both of them were applied on the established subcutaneous 9 L glioma animal models and the corresponding reaction was observed. Results: Realgar showed strong inhibitory effects on the 9 L glioma cells in vitro. According to the trypan blue coloration and cell counting, the suppression ratio of the cells on 4 h, 12 h and 24 h was 40%, 78% and 99% respectively; in the animal model experiment, strong antitumor effects were also observed. In the local injection group, about 75% of the tumours disappeared in less than 3 weeks after injection， and about 8.3% of the tumours relapsed in 3 months. Conclusion: Realgar has strong antitumor effects both in vitro and in vivo,and it is a potential antiglioma agent.