Table of Content

30 April 2008, Volume 28 Issue 4

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Establishment of Postoperative Metastatic Animal Model of Human Glioblastoma in Nude Mice

YAN Ming-xia,LIU Lei,JIA De-shui,WU Hai-yan,WANG Xiao-min,KONG Han-wei,YAO Ming

2008, 28(4):  204-208. 

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Objective To establish a metastatic animal model of human glioblastoma by surgical tumor resection in nude mice, so as to provide an ideal animal model for experimental and clinical studies. Methods Histological intact tumor lesions of human glioblastoma U87MG were transplanted subcuta-neously into twenty-seven nude mice. When the tumor reached 20 mm in diameter, eighteen of these nude mice were randomly selected as experimental group and had their tumors resected to establish the postoperative metastatic animal model. The rest were assigned to the control group. All mice were sacrificed when the animals were obviously slender，the tumor metastasis rates and pathological changes were observed. Results The postoperative metastatic animal model of human glioblastoma were successfully established. In the experimental group，the average survival time of nude mice was 67.3 days, the average lung weight was 0.83士0.30 g，the metastasis rates in lung, lymph node, and cardia was 100% (18/18)，66.67% (12/18) and 5.56%(1/18), respectively. While in the control group, the average survival time of nude mice was 56 days，the metastasis rate of lung was 11.11% (1/9)，the average lung weight was 0.41±0.12 g. Conclusions This animal model simulates the metastasis after clinical operation, it is an ideal animal model for pathogenesis exploration and anti-metastasis treatment of human glioblastoma.

Comparison of Heart Function Changes by Transverse Aortic Constriction in Different Strain Mice

LI Xiao-mei1,MA Yi-tong1,YANG Yi-ning1,LIU Fen1,Huang Ying1,HAN Wei1,GAO Xiao-ming2,ZHANG Jian-fa1

2008, 28(4):  209-214. 

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Objective To compare difference of the function of the hypertrophic hearts induced by transverse aortic constriction(TAC) in different strain mouse with microsurgery. Methods KM and C57BL/6 mice randomly divided into three groups: before operation (BO)(n=10)、sham-operation (SH) (n=10) and transverse aortic constriction(TAC)(n=30). Echocardiographic assessments, hemodynamic determineation, organ weight measurement and histological examination were performed before and at 8 weeks after surgery, and observed the cause of death. Results The successful rate of KM and C57BL/6 TAC groups was 80% and 87%. The survival rate of two TAC groups was 67% and 77% after 8 week; Compared respectively with SH group, at 8 week after surgery, anterior wall thickness at systole and diastole (Awsth; Awdth)、posterior wall thickness at systole and diastole (Pwsth; Pwdth)、LV mass index (LVMI)、systolic blood pressure(SBP)，diastolic blood pressure (DBP)、left ventricular systolic pressure (LVSP)、left ventricular end-diastolic pressure (LVEDP)、rate of rise of left ventricular pressure (士 dp/dtmax)、the heart weight-to-body weight ratio (Hw/Bw)、left ventricular Weight(LVw)、the LV weight-to-body weight ratio (LVw/Bw) of KM and C57BL/6 TAC groups increased significantly(P＜0.05). Compared with KM SH group, the left ventricular end-systolic dimension (LVESD)、Ventricular end-diastolic dimension (LVEDd), external left ventricular diastolic diameter (EXLVDd) of KM TAC group increased markedly (P＜0.05). Compared with C57BL/6 TAC group, LVESD、LVEDd、EXLVDd、LVw、the Lung weight-to-body weight ratio(Lungw/Bw) of KM TAC group increased markedly (P＜0.05).Conclusion KM mice can establish a TAC model that induces the myocardium of LV from hypertrophy tc heart failure. The result is as the same as C57BL/6 mice’s. KM mice shows the tendency of eccentric hypertrophy early.

Clone and Identification of β-defensin Gene in Mongolian Gerbil

LIU Yue-huan1,WU Jiu-sheng2,SHI Zhang-Kui1

2008, 28(4):  215-219. 

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Objective To Clone p-defensin gene in Mongolian gerbil. Methods The primers was designed according to the published sequences in GenBank. A RT-PCR method was applied to clone the defens in beta 1 gene in small intestine of the Mongolian gerbil. The PCR products was sequenced and blasted with related sequences. Results The sequenced results presented that the product was highly identical to the defens in beta 1 gene in rat and mouse. The identification between the present sequence and that of rat or mouse was ranged from 85% to 78%，the GenBank Accession Number is EU784838. Conclusions The acquired sequence was a portion of defens in beta 1 gene, which presented highly identical to the same gene in rat and mouse.

Cloning and Sequence Analysis of the Flagellin B Gene of Helicobacter hepaticus

GAO Zheng-qin1,ZHANG Qiang2,HE Zheng-ming1,XING Jin1,YUE Bing-fe1,FENG Yue-fang1

2008, 28(4):  220-224. 

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Objective The flagellin B (fla B) gene of Helicobacter hepaticus (H. hepaticus) was amplified, cloned and sequenced in order to investigate its function. Method A pair of primer was used for amplifying H. hepaticus fla B gene of BJ 0801 strain with PCR. The amplified fragment was cloned into the pGEM-T vector and transformed into competence E. coli DH5a. The recombinant plasmid was identified by PCR, endonuclease analysis and electrophoresis, and confirmed by sequencing. The nucleotide sequence was compared with the fla B gene of H. hepaticus ATCC 51449. Results The results demonstrated that nucleotide homology of H. hepaticus BJ 0801 fla B gene with H. hepaticus ATCC 51449 fla B gene was 99%. Conclusion Characterization of the H. hepaticus BJ 0801 strain at the genetic identification would be an important step toward investigating H. hepaticus fla B gene function.

Morphological Characters and Genetic Polymorphism Analysis by Microsatellite Loci in Rhesus Monkey Stock from Wannan Mountains

ZHANG Hui1,2,LI Jin-hua1,2,ZHAO Jian-yuan1,3,WANG Teng-hai4

2008, 28(4):  225-229. 

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Objective In order to establish the closed colony, the heredity background of the rhesus monkeys from Anhui stock were investigated. Method Several morphological variables of 18 individual monkeys from Anhui Wannan Mountains were measured, and the genetic polymorphism of 33 monkeys was analyzed by micro satellite loci. Result The results of f-test showed that there were evident differences in weight between male and female and that in trunk length between adult and subadult. At the same time, nine polymorphic microsatellite loci were selected from twelve loci. The mean number of alleles of Anhui rhesus monkey stock detected was 8. The heterozygosity value was 0.825±0.106, and the polymorphism information content value was 0.788±0.121. Conclusion Morphological traits of Anhui rhesus monkey stock embodied the feature of M.mulatta littoralis. And genetic diversity of the group is high on the microsatellite level. So we can screen out the original seed from Anhui rhesus monkey stock and establish the closed colony.

Influence of Dexamethasone on Blood TNF-α,IL-10,GR in Rats Models of Acute Necrotizing Pancreatitis

LIU Na-xin1,LIU Cun-li2,CHEN Zhou-Xun1,SHI Hong-Qi1

2008, 28(4):  230-233. 

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Objective To investigate the changes of TNF-a, IL-10, glucocorticoid receptor (GR) in Rats with acute necrotizing pancreatitis (ANP) and the influence of Dexamethasone(DEX) on these parameters in ANP rats. Methods Fourty-eight male Sprague-Dawley (SD) rats were randomly divided into acute-necrotizing-pancreatitis group (group A,n=16)，DEX-treated group (group D, n=16) and sham-operation group(group S，n=16). ANP model was induced by intergraded injection of 5% sodium tanrocholate to biliopancreatic duct in the rats except for group S, and the rats in group D were intramuscularly given dexamethasone after operation. The change of pancreatic pathology, the serum AMY, TNF-a and IL-10 level, and the blood mRNA expression of GR were examined. Results ①Typical ANP pathological changes were found in group A, including i.e. interstitial edema, infiltration of inflammatory cells, hemorrhage and parenchyma necrosis; and the mild inflammation was found in rats of D group which treated by Dexamethasone.② The concentration of serum TNF-a, IL-10 in group A were obviously higher than that in group S (P＜0.01). The concentration of serum TKF-a decreasing and IL-10 increasing in rats were induced by the usage of Dexamethasone (P＜0.01).③ The mRNA expression of blood GR in group D was lower significantly at 8h or 12h after the operation than that in group S (P＜0.01 or P＜0.05). Conclusions The changes of TKF-a, IL-10, GR are involved in the inflammation response of rat ANP, and the dexamethasone can relieve ANP inflammation response by affecting these factors.

Selection of Organic Cosolvents in Experiment with Zebrafish

WANG Si-feng, WANG Xi-mm，HOU Hai-rong, LIU Ke-chun, HAN Li-wen, CHEN Xi-qiang, WANG Xue

2008, 28(4):  238-242. 

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Objective To study the applicability of four kinds of organic solvents, dimethyl sulfoxide, acetone, methanol, isopropyl, as cosovlents in anti-tumor drug screening with zebrafish model. Methods zebrafish embryos at the stage of 12h or 24 h post-fertilization were treated with serial concentrations of each organic solvent. The state of angiogenesis，abnormality and death of the above zebrafish embryos were investigated by counting the number of intersegmental vessels and calculating rates of abnormality and death until 72h post-fertilization. Results The shape and number of intersegmental vessels of living zebrafish embryos in each organic solvent are normal and identical with the control. Abnormality and death of zebrafish embryos are not observed in 0.1% concentrations of dimethyl sulfoxide, acetone and 1 % concentrations of methanol，while rates of abnormality and death are significantly higher than the control with the increase of concentration. Abnormality and death rates of zebrafish embryos are significantly higher in 0.1% concentrations of isopropyl compared with the control. Conclusions Used as a cosolvent, the suitable concentration for dimethyl sulfoxide and acetone is below or equal to 0.1 %, and for methanol is below or equal to 1 %. Isopropyl is not fit for anti-angiogenesis drug screening with zebrafish model.

A new method to make water for the Morris water maze

LIU Yan-xia1，HU Jm-hong1，ZHU Quan-gang1，LIU Ji-yong, ZHENG Jie-min2

2008, 28(4):  243-245. 

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Objective To develop an economical and practical method by adjusting water color and turbidity for Morris water maze Methods Pigment was used to adjust the color and turbidity of the water by screening the suitable concentration and proportion. Results 10 mg/L white pigment was suitable for the requirenent of turbidity, 5 mg/L melanin for color, and the mixture of 10 mg/L white pigment and 5 mg/L melanin for the turbidity and color perfectly. Conclusion 10 mg/L white pigment could be used for Morris water maze tests in small pigmented rodents and the mixture of 10 mg/L white pigment and 5 mg/L melanin solution was suitable in the small albino rodents.

Experimental Study on Establishment of Encephalitozoonosis Model with FMMU Albino Guinea-pig

WANG Wan-shan1, QU Li-zhi1, LIU Zhen-xi2, WU Qing-hong1, Zhang Jia-ning1, GU Wei-wang1

2008, 28(4):  246-250. 

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Objective To establish encephalitozoonosis model of FMMU albino guinea-pig. Method FMMU albino guinea-pigs were divided into 2 groups: immunosuppressed group, non-immunosuppressed control group. Encephalitozoon cuniculi were isolated and purified from diseased rabbit, guinea-pigs were infected by intragas trie administration of E. cuniculi, and hydrocortisone was administered by intramuscular injection. E. cuniculi in faeces，clinical manifestations, and histopathologic changes in major organs were observed, the ultrastructure of E. cuniculi was observed by electron microscope(EM). Result E. cuniculi were detected from hydrocortisone-immunosuppressed guinea-pigs at day 3 post infection, one day earlier than that of non-immunosuppressed ones, and all immunosuppressed guinea-pigs were detected with E. cuniculi at day 6, ahead of control group 2 days. Minority of guinea-pigs in control group showed loss of appetite and emaciation, others were asymptomatic. Emaciation and ascites could be seen in immunosuppressed guinea-pigs, and 5 of which had neurological and urinary symptoms such as wryneck, convulsion, oliguria and painful urination at day 35 post infection. Mortality rate was 25% at day 40 post infection in immunosuppressed group. Under light microscope there were focal lymphoepithelioid granuloma in internal organ, such as brain, kidney and liver. EM observation showed that E. cuniculi spore has a trilaminar wall, the interior polar tube was arranged into 6 coils. Conclusion E. cunicul infection model of immunosuppressed FMMU albino guinea-pig was established successfully and could be used as acute Encephalitozoonosis animal model for human and other animals.

Changes and Significances of Apelin and its Receptor in Kidney Tissue of Rats Chronically Exposed to Hypoxia

ZHOU Zhu-ying, MAO Sun-zhong, GONG Yong-sheng，FAN Xiao-fang, TIAN Li-xian, Huang Ping

2008, 28(4):  251-254. 

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Objective To study the expression changes of apelin and its receptor APJ at kidney tissue of rats chronically exposed to hypoxia. Methods Twenty male Sprague-Dawley rats were randomly divided into control group and 4-week hypoxia group. The levels of blood urea nitrogen (BUN), serum creatinine (Scr) and renal tissue hydroxyproline were measured. The protein level of apelin in the kidney was measured by radioimmunoassay. The mRNA expressions of apelin and APJ in kidney tissue were determined by reverse transcription polymerase chain reaction. Results ① The content of hydroxyproline in hypoxia group was 37.31%(P＜0.01) higher than that in control group ,while there were no significant differences of Scr and BUN between the two groups.② The expressions of apelin and APJ mRNA in kidney tissue of hypoxia group were higher than those of control group (P＜0.0).③ The protein level of apelin in kidney of hypoxia rats also was obviously higher than that of control ones. Conclusion The increased protein and gene expressions of apelin and APJ in kidney tissue of rats chronically exposed to hypoxia might be an important factor in the development of renal interstitial fibrosis.