Table of Content

25 August 2011, Volume 31 Issue 4

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Comparative study on Liver Fibrosis Models Using uPA Knock-out Mice and Wild-type Mice

WANG Xiao-dong, ZHANG Yan, WANG Yu-zhu, JIN Jia-jing, CHENG Qian-qian, LI Wei-hua, DING Xun-cheng

2011, 31(4):  231-235.  DOI: 10.3969/j.issn.1674-5817.2011.04.001

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Objective To build and compare two liver fibrosis models using uPA knock-out (uPA-/-) mice and wild-type (WT) mice. Methods Adult male C57BL/6J WT mice and uPA knock-out (uPA-/-) mice were divided into four groups with 10 mice in each group: control groups (WT, uPA-/-) and liver fibrosis model groups (WT, uPA-/-). Mice were injected intraperitoneally with 0.15ml 10% CCl₄ (or olive oil as control) 2 times per week for 4 weeks and 6 weeks respectively. Serum alanine aminotransferase (ALT), aspartate(AST), albumin(ALB) and albumin/globulin(A/G) were determined, liver hydroxyproline (Hyp) level was determined using hydrochloric acid hydrolysis method and serum procollagen III (PC III) content was measured by radioimmunoassay; liver histopathology were performed using HE staining and Masson staining. Results After modeling for 4 weeks and 6 weeks, serum ALT and AST activity of both WT mice and uPA-/- mice significantly increased while serum Alb content significantly decreased comparing with control (P<0.01); at 4 weeks, significant decrease of A/G content was found in uPA-/- mice model only (P<0.01). After modeling for 4 weeks and 6 weeks, serum PCⅢ level and liver Hyp content of uPA-/- mice model significantly increased comparing with control (P<0.01); At 6 weeks, serum PCⅢ level and liver Hyp content of WT mice models started to increase significantly (P<0.01), and the elevated values were not as high as those in uPA-/- mice model. Histopathology showed that the degree of liver fibrosis in uPA-/- mice model was more severe than that in WT mice model. Conclusion Compared with C57BL/6J WT mice, establishment of liver fibrosis model by using uPA-/- mice was required shorter time and showed more severe degree of liver fibrosis.



Evaluation of Exercise Therapy's Effect on Rats' MACO Model by Functional Magnetic Resonance Imaging (fMRI) Combined with Nervous' Symptom Score

ZENG Gui-gang, ZHANG Shen, CHEN Guo-qiang, GU Jian-zong, GU Wei-zong

2011, 31(4):  236-241.  DOI: 10.3969/j.issn.1674-5817.2011.04.002

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Objective To evaluate the effect on the stoke model of rat by functional magnetic resonance imaging; evaluation (fMRI) combined with the nervous' symptom score. Method After successfully modeled, 40 MACO model rats were randomly divided into model group and exercise group, 20 animals were assigned in each group. The recovery of the rats was synthetically evaluated by the nervous symptom score on Day1, 10, 20, 30. The cerebral infraction position in both groups was detected by fMRI on Day3, 30. Result The score in both groups on Day30 was significantly higher than the Day1. But the score in exercise group's was significantly higher than the model group (P<0.05). The T1WI, T2WI were normal, and diffusion ensor ractograph (DTT) showed bilateral nerve fiber bundle was symmetrical without interruption in normal rats. On Day 3, the exercise group, the T1WI showed low signal strip shadow and the T2WI showed large flake of high signal shadow in right cerebral hemisphere, the fiber bundle in right hemisphere was significantly decreased and broke off compared with the left hemisphere. On Day 30, T1WI, T2WI showed large areas of malacia in right hemisphere, the fiber bundle was obviously fewer than the opposite side and have been broken off. The blood oxygenation level dependent-functional magnetic resonance imaging (BOLD-fMRI) discovered the activated spot appeared in very different position and had poor reproducibility in different group rats and before-after study in the same rat. Conclusion The exercise therapy method could significantly improve the nervous symptom of stroke model rats. DTT could accurately detect the pathologic change of the nerve fiber bundle after the rats have been stroked. BOLD-fMRI was not appropriate to detect the rat's allokinetic activation position under anesthesia state.



Establishment of Mice Model of Lung Cancer with Lung and Kidney Deficiency

WANG Ju-yong, GUO Jing, ZHENG Zhan, WANG Qing

2011, 31(4):  242-247.  DOI: 10.3969/j.issn.1674-5817.2011.04.003

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Objective To establish a mice model of lung cancer with lung and kidney deficiency. Methods A total of 24 eight-week-old male C57BL/6 mice were randomly divided into normal group, Lewis lung cancer model group (model group)and Lewis lung cancer with lung and kidney deficiency group (combined group),with 8 mice in each group. Mice in normal group received no treatment, mice in model group were injected subcutaneously to the right axilla with Lewis lung cancer cells. The combined group were injected with Lewis lung cancer cells and hydrocortisione (25mg/kg, 1 times/day, × 14 days) and stimulated with cigarettes (2 times/days, × 14 days).The model of lung cancer with lung and kidney deficiency was determined by observing behaviors and physical signs of the mice, detecting the changes of feed consumption, body weight, survival time, tumor weight, spleen index, thymus index, lung index and histopathology. The changes of number of CD4⁺CD25⁺Foxp3⁺Treg cells and the expression of Foxp3 mRNA in spleen, thymus and tumor were detected by flow cytometer and real-time PCR. Results Compared with those in the normal group and model group, the mice in the combined group were noted with obvious signs of lung and kidney deficiency, such as activity less, sleepless, curly, hair loss, tiredness, obvious weight loss, rapid shallow breathing and urinating more than the amount of the performance, et al. Especially, the median survival time in combined group was significantly reduced 19 (13-33) d (P<0.01). The number of CD4⁺CD25⁺Foxp3⁺Treg cells in spleen and thymus was higher in combined group than in normal group and model group (P<0.01). The expression of Foxp3 mRNA in spleen and thymus was significantly up-regulated, compared with those in combined group than in normal group and model group (P<0.01). Conclusion The mice model of lung cancer with lung and kidney deficiency is established. It is provided an experimental tool for the study the mechanism Traditional Chinese Medicine treatment with lung cancer.



Pathological Changes of Multi-organ in Acute Respiratory Distress Syndrome Induced by Lipopolysaccharide in Sheep

NIU Ting-xian, LUO Xiao-hong, WAN Dong-jun, SHI Zhi-yong, WANG Hong-yi, LU Lu

2011, 31(4):  248-251.  DOI: 10.3969/j.issn.1674-5817.2011.04.004

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Objective To observe the pathological changes of multi-organ in acute respiratory distress syndrome(ARDS). Methods The sheep model of ARDS was established by infusing low dose lipopolysaccharide (LPS) intravenously. The sheep were sacrifised after 72 hours, the organs were collected and fixed by 10% formalin. Routine olefin cut sheets were made and stained by hematine-eosin. Then their pathology changes were observed under light microscope. Results Each organ has pathology change in different degree. The most severe organs of pathology change were lung and kidney, liver and spleen were secondary, but heart was not obvious. Moreover the key features of pathology change in organ were stagnant blood, degeneration and necrosis. Conclusion The multi-organ had pathology changes in different degree in ARDS by LPS inducement and lung was the first target organ that was the easiest impaired.



The Effect of Zinc on Content of Metallothionein and Ultrastructure in Liver and Brain of Rats Injected with Methylmercury

SHI Yong, WANG Zi-wei, SONG Xiang-fu, SUN Bing-xue

2011, 31(4):  252-254.  DOI: 10.3969/j.issn.1674-5817.2011.04.005

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Objective To study the effect of zinc on metallothionein content and cell ultrastructural in liver and brain of rats treated with methylmercury. Methods The rats were randomly divided into four groups: zinc sulfate group, Zn sulfate+MeHg group, MeHg group and control group. The metallothionein (MT) contents and cell ultrastructure in liver and brain were examed. Results Zinc can enhance metallothionein content in the liver and brain of rats exposed to methylmercury. The result of live and brain cells ultrastructure of rats exposed to methylmercury also reveal that cell gap widened, dense mitochondria in the cytoplasm, ribosome increased, accompanied by swelling of the endoplasmic reticulum vacuolization, chromatin aggregation. In contrast, compared with the control group. the zinc sulfate + methylmercury group is no significant difference. Conclusion Zinc can inhibit the cell damage caused by methylmercury.



Observations on Distribution of Quantum Dots and Acute Toxicity in Rats

SUN Peng, LI Peng, ZHANG Xu-rui, QIAO Wei-wei, ZHAO Xia, ZHANG Yun-peng

2011, 31(4):  255-258.  DOI: 10.3969/j.issn.1674-5817.2011.04.006

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Objective To study the acute toxicity and the distribution of quantum dots in rats. Methods Six rats underwent quantum dots injection at the subcutaneous of upper limbs while the other six rats underwent physiological saline injection at the same sites. Two weeks observation and post-observation dissection were done on all of the twelve rats to confirm if there were any abnormities. The distribution of quantum dots was measured by inductively coupled plasma-mass spectrometry. Results No apparent toxicity was seen during two weeks' observation. The results of Cadmium assay showed that quantum dots mainly accumulated in the livers, spleens and kidneys of the rats. Conclusions The toxicity of quantum dots can be affected by many factors, which remain further research.



Acute Toxicity of Oil Dispersant GM-2 to Four Species of Aquatic Animals

WANG Ying, SUN Li-ping, WEI She-lin, CHENG lin, HUANG Ren

2011, 31(4):  259-263.  DOI: 10.3969/j.issn.1674-5817.2011.04.007

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Objective To compare and determine the acute toxicity of oil dispersant GM-2 to the larvae of four species of aquatic animals and to provide reference for application and management of oil dispersant. Methods Static toxicitiy tests of oil dispersant GM-2 to Moina monggollica larvae, Ctenogobius gymnauchen larvae, Artemia nauplii and Litopenaeus vannamei postlarvae were carried out, and the measurement of 24h-median lethal concentrations (24hLC₅₀) were done. Results The 24h LC₅₀ values of the four aquatic animals were respectively 53.08, 228.98, 4132.79 and 9404.08 mg/L. Conclusion There were significant differences in toxicities of the oil dispersant, the sensitivities of the four aquatic animals to oil dispersant were from high to low Moina monggollica larvae, Ctenogobius gymnauchen larvae, Artemia nauplii and Litopenaeus vannamei postlarvae. There was strong acute toxicity of the oil dispersant to Moina monggollica larvae and Ctenogobius gymnauchen larvae, the 24 h LC₅₀ were muchlower than that of the state standard (GB 181881.1-2000).



Observation and Comparasion on Cardiac Hemodynamics in WHBE Rabbit with Acute Myocardial Ischemia

LU Ting, PAN Yong-ming, CHEN Min-li, CHEN Liang, XU Jian-qin, HE Huan, Wang De-jun, YING Hua-zhong

2011, 31(4):  264-268.  DOI: 10.3969/j.issn.1674-5817.2011.04.008

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Objective To observe the change of cardiac hemodynamics in WHBE rabbit acute myocardial ischemia model induced by ligation of coronary artery, and compare it with Japanese white rabbit to investigate the difference of the cardiac hemodynamics. Methods Male WHBE rabbits and Japanese white rabbits were randomly divided into sham operation group and model group ( =10 in each). The rabbits sham operation group were only opened chest but not ligated, and the myocardial infarction model was induced by ligation of coronary artery. The cardiac hemodynamics was observed. Results LVSP, LVDP, Lo, -dp/dt_max and RPP in normal anesthetized WHBE rabbits were significantly higher than those of Japanese white rabbits (P<0.05, P<0.01), but HR, DAP, MAP, +dp/dt_max, LVEDP and Tau has no significant difference (P>0.05).Compared with sham operation group, the change rates of SAP, DAP, MAP, LVSP, +dp/dt_max, -dp/dt_max, LVDP, Lo, RPP in WHBE rabbits and Japanese white rabbits model groups after ligation of coronary artery were significantly lower than those of sham operation groups (P<0.05, P<0.01), while the change rates of LVEDP and Tau significantly increase (P<0.05,P<0.01), and HR was trend to decrease (P>0.05). The cardiac hemodynamics parameters in WHBE rabbits after ligated for 60 min were slightly lower than those of Japanese white rabbits (P>0.05). Conclusion WHBE rabbit has a significant hemodynamics changes after ligated coronary artery, this model can be used in evaluation of anti-myocardial ischemia drug and mechanism research.



Detection of Relative Expression of ompT Gene in Escherichia coli by SYBR Green I RT-PCR

XU Xiao-jing, YIN Wei-min, ZHAO Li-xiang

2011, 31(4):  269-272.  DOI: 10.3969/j.issn.1674-5817.2011.04.009

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Objective To establish a method to relatively quantify the expression of ompT and the gene of APEC strain E058 in vivo. Methods Two-step real time quantitative RT-PCRs (qRT-PCR) of ompT and gapA were developed based on SYBR Green I, respectively. In these qRT-PCRs, ompT was identified as the target gene and gapA as internal reference, and two standard curves were established using a series dilution of cDNA synthesized from the RNA of APEC E058 grown statically to exponential phase in rich medium. Results qRT-PCR of ompT was established. In chicken challenge model, the expressions of ompT in APEC E058 were up-regulated 6.69-fold compared to that of APEC E058 grown statically to exponential phase in rich medium. Conclusion The established qRT-PCR was reliable for the expression analysis of ompT.



Protective Effects on Target-organ of Bu Yang Huan Wu Tang on SHR/SP

WU Yong-jie, WU Zhuo-qun, GU Wen-yong, GE Jian

2011, 31(4):  273-275.  DOI: 10.3969/j.issn.1674-5817.2011.04.010

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Objective To investigate whether Bu Yang Huan Wu Tang has any effect on target-organ protective effects of in stroke prone spontaneously hypertensive rats (SHR/SP), Methods Twenty male stroke-prone spontaneously hypertensive rats (SHR/SP) at 6 weeks-old were divided into 2 groups, one group rats were fed with Bu Yang Huan Wu Tang, another group rats were used as the control group. Meanwhile, 10 WKY rats with the same age were used as the normal blood pressure group.The test lasted for 3 months, the blood pressure, plasma NO, ET-1 content, rat heart, brain and kidney damage were observed. Results The damages of the target organs (heart, brain, kidney) in control group was significantly higher than treatment group and the WKY group (P<0.01). Blood pressure of the control group was significantly higher than treatment group (P<0.01) and WKY group (P<0.01). Plasma NO of control group was lower than treatment group (P<0.01). Plasma ET-1 of Control group was higher than treatment group (P<0.01). Conclusion Bu Yang Huan Wu Tang can reduce the SHR/SP blood pressure, reduce the SHR/SP target-organ (heart, brain, kidney) damage.

Effects of Hyperhomocysteinemia (HHcy) on Depression in Rats

ZHANG Xiao-feng, WANG Guo-hua, GUAN Yu, XIA Jing

2011, 31(4):  276-279.  DOI: 10.3969/j.issn.1674-5817.2011.04.011

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Objective To study the effects of hyperhomocysteinemia on depression and atherosclerosis in rats. Method Twenty male Wistar rats were randomly divided into model group and normal group (each n=10). The rat model was established by feeding L-methionine with gastric irrigation daily. After four, six and eight weeks, the total homocysteine (THcy) concentration in plasma and the immobility time of “forced-swimming test” in five minutes were respectively detected to reflect the situation of depression caused by HHcy. Eight weeks later, the change of atherosclerosis in lipid plaque coverage was detected in abdominal aorta intima. Result After six week's dosing, the concentrations in plasma of the model group (15.36±3.6 μmoL/L) were significantly higher than those in the normal group (11.38±1.78 μmoL/L, P<0.01). All the immobility times in 4 times “forced-swimming test” were prolonged and more significantly increasing (193.8±27.7 s) than the result of normal group (113.1±27.39 s) at end of sixth week (P<0.001). At same time, the percentage of lipid plaque coverage in abdominal aorta intima was significantly increased at the end of the test (the model group was 7.45%±3.99%, the normal group 4.05%±2.22%, P<0.05). Conclusion The plasma THcy concentration of model group was higher than that of normal group. The rats that had hyperhomocysteinemia (HHcy) were showed the depression and the atherosclerosis to a visible extent.



Effects of Aqueous Extract from Selaginella uncinata on Asthmatic Rats

YU Bing

2011, 31(4):  280-282.  DOI: 10.3969/j.issn.1674-5817.2011.04.012

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Objective To investigate effects of aqueous extract from Selaginella uncinata on the level of IgE in serum and IL-5 in lung of the asthmatic rats. Method Sixty male SD rats were randomly divided into six groups. Then the asthma rats model were made by ovalbumin (OVA)-sensitized and stimulated method. The level of IgE and IL-5 were detected by enzyme-linked immunosorbent assay (ELISA). Result Compared with the normal control group, the levels of IgE and IL-5 in the asthmatic group were higher (P<0.01). The aqueous extract of Selaginella uncinata could decrease levels of IgE and IL-5. Conclusion The extract of Selaginella uncinata could effectively decrease IgE and IL-5 levels on asthma rats and improve the asthmatic symptoms.

Effect of Squalene on Hematological Indices in MSG-Obese Rats

ZHANG Jin-ming, LU Hao, ZHANG wei-ming

2011, 31(4):  283-286.  DOI: 10.3969/j.issn.1674-5817.2011.04.013

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Objective To evaluate the effect of squalene on body fat and organ weight in male obese rats treated with MSG (Monosodium glutamate). Methods Thirty male Wistar rat were randomly divided into 2 groups,the model of the obesity(n=20) and the control group (n=10) at equivalent volume of saline during the same period. After weaning, the trial groups fed with squalene-containing (1000 mg/kg) diet orally and model group given a regular control diet as the same as negative control group. All rats were euthanized at 22 nd week. The concentration of blood fat and blood serum index were measured. Results Compared with the model control group the blood glucose, cholesterol, triglyceride of the squalene treated group rat were significant lower (P<0.05), LDL-C and HDL-C were lower but no significant differece (P>0.05), and Leptin and testosterone were higher (P<0.05). Conclutions The squalene may counteract increases in body fat, reduce blood glucose, cholesterol, triglyceride, LDL-C and HDL-C, increase Leptin and testosterone in MSG -obese rats.



Establishment of Liver Puncture Method in Marmota himalayan

FAN Wei, WANG Zhong-dong, TAO Yuan-qing, FENG Xue-mei, WANG Bao-ju

2011, 31(4):  287-289.  DOI: 10.3969/j.issn.1674-5817.2011.04.014

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Objective To establish the method of liver puncture in marmota himalayan. Method The animal was anesthetized by ketamine hydrochloride (45mg/kg) intramuscular injection. Liver puncture point was on the right side of the abdomen in the third and fourth ribs of animals used by biopsy puncture gun, below the sternum xiphoid 1.5 cm, with parallel the midpoint of axillary. Twenty marmota himalayan were punctured with the interval l5days, by total of eighty times from 2009 to 2010. Result Seventy-two liver tissue samples were collected and the success rate was 90%. HE staining confirmed that 70 cases of biopsy tissue had at least three portal areas in the microscope. There were two marmots died caused by liver biopsy and the death rate was 10%. Conclusion Successfully established the liver puncture method of marmota himalayan animal models of hepatitis B, the method was simple, accurate, repeated sampling.



Progresses on Application of African Green Monkeys in AIDS Researches

QIU Zheng-liang, YANG Gui-Bo, LEI Na, SUN Zao-Zheng, ZENG Ling

2011, 31(4):  308-312.  DOI: 10.3969/j.issn.1674-5817.2011.04.021

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Human infected with human immunodeficiency virus (HIV-1) always progress to AIDS after a variable period of latency. However, African green monkeys (AGM) with high levels of virus do not show any signs of AIDS after natural infection with simian immunodeficiency virus (SIV). This elicited interest of researchers to use AGM in studies on the cellular and molecular mechanisms to control virus replication and disease progression. In this paper, recent progresses in this field were briefly reviewed.