Table of Content

  

24 July 2006
Volume 44 Issue 7

Articles


Articles

Cloning of human NKp46 gene and its expression and purification in E.coli

GAO Xin-pu,LIU Zheng-min,WANG Lai-cheng,JIAO Yu-lian,LIUYi-qing,ZHANG Xue,ZHAO Yue-ran

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(7):  649-653. 

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To clone the gene of human NKp46，express and purify the recombinant human NKp46 in E.coli. Methods：A hNKp46 DNA fragment，with a length of about 900?bp，was amplified from the total RNA of peripheral blood mononuclear cells by RTPCR and cloned to plasmid pMD18T，and then the cloned DNA fragment was sequenced.then hNKp46 fragment was isolated and inserted to the corresponding restriction site on procaryotic expression vector pET30a(+). The recombinant plasmid pET30a(+)hNKp46 was identified by enzymogram and transformed to E.coli BL21(DE3)，and then its expression was induced by IPTG. The expressed product was identified by SDSPAGE and Western Blotting, and the expressed protein was purified by His·Bind Purification Kit. Results：The length of DNA fragment amplified by RTPCR was consistent with that of hNKp46 cDNA. DNA sequencing of pMD18ThNKp46 revealed that the cloned DNA sequence was identical to that of reported hNKp46 cDNA. SDSPAGE proved that expressed product， with a relative molecular weight of 38.5?kD，contained about 40% of total somatic protein. Western Blotting showed that the recombinant protein could specifically bind to antiHis·Tag antibody. The recombinant protein was obtained by purification with 95.5% final purity and 40% recovery rate. Conclusion：A recombinant bacterial strain for expressing hNKp46 is successfully constructed, and its recombinant protein is purified.

Effect of monastrol on mitosis in human pulmonaryadenocarcinoma cell line A549

ZHAO Jian,CHEN Wei-wen,WANG Shu-cheng,LI Xi-bo,

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(7):  658-661. 

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To explore the influence of monastrol on mitosis in human pulmonary adenocarcinoma cell line A549. Method: Immunofluorescence stainning and fluorescenceactivated cell sorting techniques were used to detect the the count of cells and the cell cycle. Result: A549 cell was blocked in prometaphase by monastrol and the mitosis cells were monopolar spindlelike. Conclusion: Monastrol can influence mitosis of lung cancer cells, and therefore inhibits the growth of lung cancer cells.

Effects of bushenhuoxue complex on cytokine and anticoagulant factors in chronic cerebral circulation insufficiency model of subacute aging rats

REN Guo-hua,JIA Qing,LI Lu-yang

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(7):  676-679. 

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To observe the effect of bushenhuoxue complex on cytokine and anticoagulant factors in chronic cerebral circulation insufficiency model of subacute rats and to investigate the mechanism. Methods: Sixty healthy male Wistar rats were randomly divided into 6 groups: common， model， highdose treatment, lowdose treatment, and control group. Subacute aging model was induced by Dgalactose, and the bilateral arteria carotis communis was ligatured. The levels of SOD, MDA, T, Fib, ATIII, ET, TNF, and Pselectin in plasma were observed. Results: The morphology of brain was ischemic changed and had no cerbral infarction in model group; The level of SOD, NOS, T, ATIII dropped off（P＜0.01） d of ET, TNF, MDA, Fib, and Pselectin rised（P＜0.01in model group，whereas the former rised（P＜0.05）and the latter dropped off (P＜0.01，P＜0.05) in therapy group; The effect for them in highdose group was superior to lowdose group except for MDA and ET(P＜0.01，P＜0.05). Conclusion: Bushenhuoxue complex can correct the disequilibrium of cytokine and anticoagulant factors, which maybe by expanding the cerebral vessel, eliminating the free radical, inhibiting the inflammation and collecting the platelet.

Renal retransplantation in 41 cases

WANG Hong-wei,LIU Shuang-de,YAN Jie-ke,TIAN Chuan,XU Dong-sheng

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(7):  722-725. 

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To summarize the longterm surval rate of the recipients receiving renal retransplantation. Methods：The clinical data of 41 cases of renal retransplantation between January 1999 and December 2005 were analyzed retrospectively. Of them, 18 underwent allograft nephrectomy (group 1) and 23 did not (group 2). The graft survival rate at 1, 3, and 5 years, the factors contributiving to the mortality and factors affecting the graft survival were observed. Results：The 1, 3, and 5year graft survival rates were 85.4％, 76.2％, and 65.8％, respectively, and those in group A and B were 88.9％ vs 82.6% (P >0.05); 74.7% vs 77.1% (P>0.05), and 62.2% vs 68.5% (P>0.05), respectively. Acute rejection (AR) rates were not significantly different between groups A (33.3%) and B (30.4%) (P>0.05). Incidence of AR was higher in patients with positive PRA, however, antibodyinducing therapy could lower the incidence of AR. Conclusions：Graft nephrectomy before retransplantation can not lengthen the longterm survival, and high quality of HLA matching and antibodyinducing therapy may improve the longterm survival rate of graft.

Effect of KAI1/CD82 and ECadherin expression in epithelialovarian carcinoma

HAN Qiu-yu,ZHANG Xiang-ning,LI Xiao-cui,CAO Li

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(7):  729-733. 

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To investigate the relationship between epithelial ovarian carcinoma（EOC）and levels of the tumor metastasis suppressor gene KAI1/CD82 and the adhesion molecule ECadherin（Ecad）. Methods：Immunohistochemistry SP method was used to examine the expression levels of KAI1 and Ecad in 66 specimens of EOC. Results：Of them, 38 showed positive KAI1/CD82 protein (57.58%) and 31 positive Ecad protein（46.97%. The statistical evaluation revealed a close relation among the clinical stage, lymph node metastasis and the expressions of KAI1/CD82 and Ecad（P<0.05）, and an inverse relationship between the histological grade and KAI1/CD82 expression levels(P<0.05), suggesting expressions of KAI1/CD82 and Ecad had great consistency in the same sample. Conclusion：Expressions of KAI1/CD82 and Ecad protein are closely related with the malignant progression and metastasis of EOC, which have important implication for evaluation of the differentiation and tumor progression and prediction of the prognosis in EOC.

Hemorheological changes of hypertension in aged patients

HUANG Ping,WANG Cui-ping,LI Xiao-guang,WANG Xian-xian

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(7):  743-745. 

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To explore the hemorheological changes of hypertension in aged patients. Method: The hemorheological changes were detected and analyzed in 178 aged patients with hypertension and 72 normal controls. Result: The levels of the hemorheological parameters(HCT, Arbc, Fib, et al) in patients were significantly higher than those in the normal controls (P<0.01). Conclusion:Aged patients with hypertension are in a state of high viscous and high coagulability, and are tending to have embolism, cardio vascular or cerebral vascular diseases, and therefore measure for reducing the whole blood viscosity should be taken.

Sequential chemotherapy of Epirubicin，Taxotere，and 5Fluorouracilincreases the apoptosis in breast cancer cell lines

RUAN Yong-wei,JIN Xing,HOU Lian-ze

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2006, 44(7):  750-753. 

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To evaluate the activity ofhe sequence exposure of Epirubicin(Epi), Taxotere(Tax), and 5Fluorouracil(5Fu) for human breast cancer so as to select the most effective treatment schedule. Methods：The cytotoxic activity of drug on MCF7 was evaluated by sulphorhodamine B assay and the interaction of the different drugs was assessed by median effect principle. The cell cycle and apoptosis were evaluated by flow cytometry. The expressions of p53, bcl2, p21,bax and thymidylate synthase(Ts) were detected by Western blotting. Results：As a single agent, 5Fu exerted a low cytotoxic activity in MCF7 cells. The Epi→Tax sequence induced a synergistic cytocidal effect and enhanced the efficacy of 5Fu. The Epi→Tax sequence blocked MCF7 cells in G2M phase, and the addition of 5Fu forced the cells to progress through cell cycle or killed them. The Epi→Tax sequence induced a significant reduction in basal Ts expression. The sequence of Epi→Tax→5Fu produced a synergistic and highly scheduledependent interaction(CI<1),