Table of Content

31 March 2010, Volume 30 Issue 3

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Comparison of Different Ischemic Methods in Establishment of Periventricular Leukomalacia Rat Model

HE Yue-qiu,CHEN Hui-jin,QIAN Long-hua,CHEN Guan-yi

2010, 30(3):  153-157. 

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Objective To compare different ischemic methods for establishing a reliable neonatal rat model for periventricular leukomalacia (PVL) which is expected to be similar to PVL in human preterm infants pathologically. Methods Two-old-day neonatal rats were randomly divided into bilateral common carotid artery ligation group，unilateral common carotid artery ligation group, and sham group.The weighing, cerebral pathological assessments under either light or electronic microscopes were respectively undertaken 2 days and 21 days after operation. Results The body weights in bilateral common carotid artery ligation group was reduced significantly compared to those in either unilateral common carotid artery ligation group or sham group (P＜0.05 for all). The relatively intact cortex but obvious pathological change of white matter in 2 days after operation and the cystic, loose, and necrotic white matter in 21 days after operation were observed under light microscope in bilateral common carotid arteries ligation group，while a diffuse neuronal injuries in cortex with a basic normal white matter was found in unilateral common carotid artery ligation group. There was significant difference in patho-logical score of white matter injury on day 21 after operation (X²=8.751, P=0.003). The myelination under electronic microscope reduced remarkably in bilateral common carotid arteries ligation group.Conclusions The ligation of the unilateral common carotid artery can induce cortical damage and the ligation of the bilateral common carotid arteries can result in white matter injury principally, and suggesting the ligation of the bilateral common carotid arteries is a reliable method for establishment of PVL model.

Effects of Curcumin on Brain Damage Repair in Cerebral Cortex and Hippocampus Tissue of Rats with Hypoxic Ischemic

YIN Guang-mei1,YU Lin-sheng2,ZHENG Yuan-yuan2,YI Ji-pu2,YE Guang-hua2,WU Shu-zhen2,ZHAO Kang2

2010, 30(3):  158-162. 

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Objective To observe the effects of curcumin on apoptosis and caspase-3 expression in hippocampus tissue area in cortex with hypoxic ischemic brain damage. Methods A total of 48 SD rats were randomly divided into four groups as the following: sham group(SH)，hypoxic ischemic brain damage group(HIBD)，curcumin group(CU) and solvent control group(SC). Brain specimens drawn from the left cerebral cortex were tested for the enzyme activity of NOS. The expression of caspase-3 protein in hippocampus tissue were detected with immunohistochemistry (IHC), ultrastructural changes of cortex tissue were observed by transmission electron microscopy. Results The The enzyme activity of NOS in CU group was clearly lower than those in HI group at the same reperfused time (P＜0.01). The expression of caspase-3 was decreased in cortex tissue in CU group than that in IR group, the number of apoptosis neurons in hippocampus tissue were much less in CU group than that in HI group.Conclusion Neuron apoptosis participates in hypoxic ischemic brain damage. The protective effects of curcumin include suppressing the expression of caspase-3 and blocking apoptosis in hypoxic ischemic brain damage.

Establishment and Change of Rat Model of Postpartum Urinary Incontinence

HUANG Jian,CHENG Ming-jun,HUA Ke-qin

2010, 30(3):  163-166. 

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Objective To observe the injury of pelvic muscle and change of pelvic function after the establishment of postpartum urinary incontinence (PPUI) animal model. Methods To establish PPUI animal model, a hyponome was placed in the rat vagina for a long time to imitate the way in which birth trauma destructs the rat pelvic floor and then sneezing experiment, urodynamic test (largest bladder capacity, leak point pressure), and the histological identification was made for observing the function of pelvic floor muscle contraction strength respectively after 1 day, 7 days 14 days, 28 days, 56 days. Results Urodynamics (largest bladder capacity, leak point pressure) and pelvic floor muscle contrac-tion strength changed in the rats of the test team. Conclusion Placing a hyponome in the rat vagina for a long time to imitate the way in which birth trauma destructs the rat pelvic floor can establish a PPUI animal model like the postpartum urinary incontinence Zs symptom in human.

Research on Bile System in Zebrafish(Brachydanio rerio)

CHU Xiao-hong1,PU Jun-yi1,2,YAO Yi-lin1,CHENG Qiu-sheng1,HU Jian-hua2,GAO Cheng2

2010, 30(3):  167-173. 

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Objective To research on the intrahepatic biliary system in the zebrafish, Brachydanio rerio. Methods By using enzyme histochemical characterization, immunohistochemistry and electron microscopy. Results The biliary system was arranged in arborization in the parenchyma.The biliary tree included sequentially by intracellular and intercellular canaliculi，preductules,ductules and ducts.The intercellular originated at the neighborhood of the nucleus to extend to the cell surface where it emptied into the intra parenchymal biliary passage running in the interhepatocytic space. In the preductules，the wall was formed by bile preductular epithelial cells and hepatocytes .The bile preductular epithelial cells were linked to hepatocytes by the tight junctions. The wall of the biliary ductule usually consisted integrally 2 to 4 biliary epithelial cells. These cells had no basal lamina and were contacted with hepato-cytes directly. A ductule that was composed of a single cell and was embedded between two adjoining liver cells was found by TEM. The lumen of this ductule was very narrow and formed by the infolding of the plasma membrane of the ductule cell. The wall of bile ducts was composed of cuboidal epithelial cells, and the bile duct was encircled by the connective tissue. Whereas in ducts, apart from those cells，there were intraepithelial lymphocytes.This suggested that there exited mucosal immune response. The preducules and ductules formed a segment connecting canaliculi to ducts, thus, we considered that segment as being analogous to the mammalian canals of Hering.The reactions for Mg²⁺-dependent ATPase showed that bile tree was strained brownish-black color and interwove into a network. The bile system was revealed by immunstaining with a monoclonal antibody 2F11 that detected biliary epithelial cell bodies which are distributed scattcrcdly in interhepatocytic and the proximal regions of the ducts arising from these cells was stained. Conclusions The bile systerm in zebtafish is not only different with the mammals and birds, but also with other bony fish.

Generation of Porcine Monocyte Derived Dendritic Cells in vitro: Morphological, Phenotypic and Functional Properties

ZHOU Qian-jun1,2,FAN Hui-min1,LIU Zhong-min1

2010, 30(3):  174-179. 

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Objective To amplify and identify dendritic cells (DC) generated in vitro from porcine peripheral blood monocyte (PBMC) and study their immunological characteristics. Methods Monocyte-derived DC (MoDC) were generated by stimulating PBMC with granulocyte-macrophage colony-stumulating factor (GM-CSF) and interleukin-4 (IL-4)，after 5 day culture，LPS (1 ug/ml) was added.Non-adherent cells were collected 5-8 days alter culture. Morphological features were characterized by phase contrast photomicrograph. The co-stimulation molecules, SLAII and other markers were assessed by FACS. Immunological functions were evaluated by means of mixed lymphocyte reaction (MLR) and ELISA. Results Porcine MoDC showed typical DC morphological features, phenotype were CDl⁺CD14⁺CD80⁺CD86⁺CD172a⁺SLA Ⅱ⁺; the expression of CD80, CD86 and SLAII elevated after LPS stimulation. Porcine MoDC showed potent allogenic T cell stimulatory capacity inducing a strong interferon-γ and IL-4 response. Conclusion Porcine MoDC were successfully generated in vitro by stimulating PBMC with GM-CSF and IL-4, that might be meaningful for the further research on tolerance induction and xenotransplantation.

Observation on Pelvic Lymph Nodes Aggregation by Cervical Injection of Fluorescent Nanoparticles in Rabbits

FENG Hua,CHENG Ming-jun,XU Cong-jian

2010, 30(3):  180-182. 

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Objective To investigate whether there is enrichment of NaYF₄ nano-particles in pelvic lymph node by cervical injection in rabbits, and also to decide the peak time and the best diameter of nano-particles. Methods 10% solution with different diameters of nano-particles (20、50、100 nm) is injected through female rabbit's cervix. The pelvic lymph nodes were be taken at different time (2、10、30、60、120 min) after injection and the aggregation peak of nano-particles were measured;inlO min 50 nm group, the liver、spleen and kidneys are taken with pelvic lymph nodes together to detect the aggregation characteristics of nano-particles. Result The 50 nm is a preferable NaYF₄ nano-particles diameter for NaYF₄ solution injection through cervix，and 10 min is the peak time.Compared with the liver，spleen and kidneys，more nano-particles were aggregated in pelvic lymph nodes. Conclusion NaYF₄ nanoparticle solution through cervical injection of the pelvic lymph nodes in rabbits can be enriched.

Optimization for Nested-PCR Method for Bacillus piliformis Detection and Its Application

FENG Jie1,2,XIE Jian-yun1,2,HU Jian-hua1,2,GAO Cheng1,2

2010, 30(3):  183-187. 

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Objective To optimize nested-PCR of Bacillus piliformis detection and to explore its application. Method The liver from rats infected by Bacillus piliformis RJ was used as the positive control. This sample was positive for Bacillus piliformis detected by indirect immunofluorescence(IFA) according to the GB/T 14926.10-2008《laboratory animal- Method for examination of Tyzzer's organism GB/T 14926.10-2008》. Nested-PCR of Bacillus piliformis established previously by our lab was optimized, including DNA extraction and PCR conditions. This optimized nested-PCR was applied to detect the Bacillus piliformis of 22 immunosuppressive rats. Results Phenol/chloroform DNA extraction method of liver was optimal; A specific 196 bp DNA fragment was obtained, the sensitivity of this method reached to 1 pg; Blast results showed that all the sequences of the specific DNA fragment shared 100% homology with the 16 S rRNA of Bacillus piliformis presented in Genbank (D14638.1).Twenty-two immunosuppressive rats were detected for Bacillus piliformis. The positive rate by IFA is 36.4%(8/22); The positive rate by nested-PCR is 59.1% (13/22indicating nested-PCR is more sensitive than IFA. Meanwhile all the positive samples detected by EFAwere also positive detected by nested PCR.Conclusion This optimized nested-PCR method has high sensitivity and specificity and can be applied to Bacillus piliformis detection.

Acute Toxicity of Melamine and Detoxification of San Jin Tang(三金汤)in Mice

ZHANG Wei1,ZHANG Zhong-guang2,XU Chong-hui3,OUYANG Shao-lun3,ZHAO Yong1,Hu Hai-xun1,LI Fu-rong1,LIN Yi-li1,ZHONG Nv-qi1,ZHUANG Shao-hui1,ZOU Yi-hai2

2010, 30(3):  188-192. 

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Objective To observe the acute toxicity of melamine and the detoxification of San Jin Tang in Kunming (KM) mice. Methods Ninety SPF KM mice (half male and female) were divided into seven groups administered with different dosage of melamine and two blank control groups. The toxic reaction to the mice caused by melamine was observed, and 70 SPF KM mice were divided into high,middle and low dosage of San Jin Tang(三金汤）administered preventive groups,San Jin Tang treatment group，melamine model group and two control groups. The detoxification of San Jin Tang in kindey and lungs after of mice administered with melamine one time was observed. Results The LD₅₀ of melamine to KM mice was 9366 mg/kg. There were no crystallization in kidney but inflammatory cells infiltration appeared m kidney, liver and lung. The content of melamine in kidneys and lung was 327ng/g and 177.5 ng/g respectively. The high dosage of San Jin Tang had best effects on cleaning up the melamine in kidney and lung, reduced to 58 ng/g and 28.2 ng/g, respectively. Conclusion The preliminary experi-ments suggested that the melamine was micro-toxic or non-toxic. The higher dose group of Chinese Medicine San Jin Tang showed best effect to get rid of melamine in mice.

Effect of Apelin on Early Renal Interstitial Fibrosis Induced by Hypoxia in Rats

WU Xiao-dong,HU Xiao-li,HONG Ling,FAN Xiao-fang,MAO Sun-zhong,GONG Yong-sheng

2010, 30(3):  193-196. 

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Objective To study the role of apelin in the prevention of renal interstitial fibrosis induced by hypoxia in rats. Methods The animal model of hypoxic renal interstitial fibrosis was established by exposing the rats to isobaric hypoxic chamber for 4 weeks (8h/d，6d/week). Thirty male Sprague-Dawley rats were randomly divided into control group, hypoxic group and hypoxia with apelin (lOnmol/kg/d) group (HA). [pGlu]apelin-13 was administered into the rats of apelin groups by mini-osmotic pump subcutaneously. Serum creatinine (Scr) and urea nitrogen (BUN) were measured. The renal homogenates were assayed for the activity of superoxide dismutase (SOD) and the contents of malondialdehyde (MDA) and hydroxyproline. Meanwhile, the renal specimens were examined by VG staining to examine renal interstitial fibrosis. Results (1) There was no significant difference of Scr and BUN among three groups. (2) The content of hydroxyproline in hypoxic group was 35.4%(P＜0.01) higher than that of control group, while apelin group was 24.6%(P＜0.05) lower than hypoxic group. (3) The activity of SOD in hypoxic group was lower than that in control group by 55.1% while the content of MDA was higher by 38.9% (all P＜0.01). Apelin treatment increased the activity of SOD by 20.7% (P＜0.01) while decreased the content of MDA by 55.7 (all P＜0.01). (4) VG staining revealed that renal vascular fibrosis was found in hypoxia groups，which was significantly alleviated by apelin treatment. Conclusions Apelin could play an important role in treatment of hypoxic renal interstitial fibrosis of rats, and the mechanisms of protection were associated with inhibition of oxidative stress.

Insulin-Resistance Tree Shrew Model Induced by High-glucose-fat-diet with Dexamethasone

LI Hai-yan,LI Jia-min,LI Jing-xiao,WANG Xin-xing,DAI Jie-jie,SUN Xiao-mei

2010, 30(3):  197-200. 

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Objective To induce an impaired glucose and lipid metabolism model by high-glucose-fat feeding (HGF) plus DEX so as to facilitate type 2 diabetes study. Methods The model was induced by HGF and HGF plus DEX respectively in tree shrew. Fourteen normal male tree shrews were randomly divided into 2 groups: HGF group with 6 tree shrews and HGF plus DEX group with 8. Several param-eters in the tree shrews were measured. Results The fasting blood glucose，lipid level and OGTT significantly increased in HGF+DEX treated group compared with the HGF group while the fasting body weight reduced in both group. Conclusions High-glucose-fat feeding and DEX together can be used to induce glycometabolism and lipid metabolism abnormality.

Interventions of Euthyrox and Stable Iodine in Hypothyroid Rats Treated with ¹³¹I

BAI Dian-qing1,LI Shi-yun2,ZHENG Ying1

2010, 30(3):  201-204. 

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Objective To investigate the effects of stable iodine and thyroxin on the hypothyroid rats treated with ¹³¹I，and provide reference in clinical ¹³¹I therapy for hyperthyroid and nontoxic goiter.Methods Fifty-four Wistar rats are equally divided into three groups，whose ¹³¹I dose was calculated by their mass. Group A accept only ¹³¹I as contrast，group B accept Euthyrox (200 pg/d, 30d) after ¹³¹I treatment, group C accept 10 )jmol stable iodine 24 h after ¹³¹I theament. Serum triiodothyronine (T₃)，thyroxine (T₄)，thyroid-stimulating hormone (TSH) are measured and cell numbers per high power lens (4 × 100) are counted in 15, 60, 90 d following ¹³¹I treatment respectively. All the data are analysed by Fisher test and q test. Results Neither cell numbers nor serum T₃，T₄ and TSH among three groups has statistically variance 15 d post treatment, With time prolonging, serum T₃，T₄ of group C decreased faster than the other two groups (in 60 d T₄ of group C is less than group B，q=5.16, P<0.05; in 90 d, T₄ of group C is less than group A than B，P<0.05). Conclusions The rats giving with euthyrox in early stage of ¹³¹I treatment can not only alleviate the damage in thyroid cells from p-rays，but also decrease the incidence of hypothyroid, and the stable iodine simultaneously probably can prolong the effective half-life of ¹³¹I，thus aggravate the damages and increase the incidence of hypothyroid.

The Dynamic Change of Some Kinds of Biochemical Indices in Serum of Rats Caused by Cold Restraint Stress

WU Jun1,REN Xiao-li2,LIN Fei-yan1,LIN Ting-ting1,BAO Jing-ying1,LOU Zhe-feng1,FANG Zhou-xi3,WU Bu-meng2

2010, 30(3):  205-208. 

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Objective To study the dynamic change of the level of ACTH、CORT、CK and ALT in serum of rats caused by cold restraint stress, and the possible mechanisms. Methods Fifty-six SD rats of 50 days old were divided randomly into 7 groups with testing time of 0 h, 0.5 h, 1.0 h, 1.5 h, 2.0 h,2.5 h，3.0 h. After putting each group rats under testing environment by using the cold restraint stress method in specified group testing time, the level of ACTH、CORT、CK、ALT and UI in serum were analyzed. Results ① The ACTH level in serum rises quickly with time increasing for the rats under cold restraint stress, it reaches the highest value at 0.5 h, and then it starts to fall quickly. (2) The CORT level rises gradually with time increasing, it reaches the highest value at 1.5 h，and then it starts to fall quickly.③ The CK and ALT levels rise gradually with time increasing for the rats under cold restraint stress.④ The UI level rises quickly with time increasing, it reaches the highest value at 2.5 h. Conclusion The cold restraint stress can cause rising of the ACTH and CORT level in serum of rat, after it reaches the highest value, it falls back quickly. It can cause CK and ALT to rise with time increasing. Cold restraint stress can cause acute gastric ulcer of rats, with the worst at 2.5 h. It suggests that the best time for creating cold restraint stress model in rat is 2.5 h.

Advances in Application of Animal Model Infected with Cryptococcus neoformans

ZHOU Wen-jiang1,2,TAO Lin-lin3

2010, 30(3):  220-224. 

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Cryptococcus neoformans is an encapsuled yeast-like fungus. It is infectious and mainly damages pulmonary tissue and central nervous system. Adult with immunodepression is prone the infection of Cryptococcus neoformans. The mortality of Cryptococcus neoformans has increased significantly in the recent decade，which caused concerns of the researchers globally. Animal models played very important roles in the systemic exploration of the mechanism of infection. Mice, rats and guinea pigs were used numerously to develop Cryptococcus neoformans infection models. Pigs, rabbits and some non-mammals were also employed as candidates to develop animal models in drug development.These kind of animal models were crucial in the research of mechanism, immunology and treatment regimen of Cryptococcus neoformans. This article is to address the current situation of development of Cryptococcus neoformans animal models, and to sum up the achievements in this field.

The Research Advancement on Animal Model of Uterine Leiomyoma

JING Hui-juan1,LI Zhen-hua2,XU Jian-yang2

2010, 30(3):  225-228. 

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Uterine leiomyoma is one of the most common benign tumors which greatly bother women who are at reproductive age. With the progress of living standard, the study of curing uterine leiomyoma draws growing attention. Establishment of animal model is a common method in research on wterine leiomyoma. This article reviewed some recent advancement on research about establishing animal model of uterine leiomyomaa.

Research Progress on Breeding and Biological Characteristics of SPF Rabbit in China

WANG Gang,LIU Ke,ZHAO Wei-jian,LI Xiong-cai,ZHANG Fu-fa,KUANG shao-song

2010, 30(3):  229-232. 

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Laboratory rabbit is an important species in laboratory animals, mainly used in pharmaceuticals,pyrogen testing of biotechnology products，the preparation of antiserum, skin irritation test，the optic nerve，microbiology，sports medicine and other scientific research and teaching fields. The requirement of laboratory rabbit is followed after the rats，mice，hamsters and guinea pigs. It is an irreplaceable experimental material and model. Research on breeding and application of SPF laboratory rabbit is rapid development in developed countries. With the development of China's life sciences and medical research，and for the request of the new “Chinese Pharmacopoeia” and “China Requirement Biological Products”，on the quality of laboratory rabbit，it was gradually moves closer to the Clean level and SPF level. This paper is an overview on the progress of the work for SPF rabbit breeding and biological characteristics in China.