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    25 December 2014, Volume 34 Issue 6
    Anatomy of Collateral Branch of Cavernous Nerves in Rat Penis and Its Effect on Penile Erection
    HU Wan-li, WANG Hao-han, YING Cheng-cheng
    2014, 34(6):  433-437.  DOI: 10.3969/j.issn.1674-5817.2014.06.001
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    Objective To explore the role of collateral branch of cavernous nerves (CN) in penile erection in rats and its importance in establishing CN injury rat models. Methods Thirty-six 3~5 months old male SD rats were used. Six rats were used to identify and observe the branches of major pelvic ganglion (MPG) under the operating microscope, the other 30 rats were divided into 3 groups randomly, including (I) sham-operated group, (II) bilateral main branch of CN ablated group and (III) bilateral main and collateral branches of CN ablated group. The erectile ability of rats were evaluated with apomorphine test respectively 7 days and 30 days after surgery. Then 4% flluoro-gold solution were injected into bilateral corpus cavernosum of penis in all rats under anesthesia, and the rats were sacrificed and their bilateral MPGs were obtained for tracing the flluoro-gold under the fluorescent microscope 5 days later. Results The main branch of CN and some fine fibers arised from the MPGs were identified on the surface of dorsolateral lobes of the prostate. 7 days after surgery, the times of rat penile erection in group was 2.0±0.7, but no penile erection was found in both group II and III. 30 days after surgery, the group II rats still showed no erection, the erectile response was appeared in group II rats after administration of apomorphine, but the erection times of group II (0.9±0.7) was still less than that of group I (2.8±0.6)(P<0.05). 5 days after injection of fluoro-gold solution, much more brighter positive stained neural cells were observed in MPGs of group II than those of group III (P<0.01). Conclusions Some tiny nerve fibers, the collateral branches of CN derived from MPG, may play a part role in penile erection of rats, and the influence of these collateral branches should be taken into consideration when creating a erectile dysfunction rat model induced by CN injury.
    Comparison of Expression Level of SLC24A5, SLC39A9 Genes in Cricetulus griseus and the Albino mutant
    LIU Hui-fang, SUN Shu-fang, ZENG Lin, CAO Yu-hua, YAN Xiang, FAN jun-wen, SUN Zhao-zeng, WANG Xin
    2014, 34(6):  438-442.  DOI: 10.3969/j.issn.1674-5817.2014.06.002
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    Objective To investigate the expression level of the soluble material transport protein SLC24A5 (solute carrier family 24, member 5, SLC24A5) and SLC39A9 (solute carrier family 39, member 9, SLC39A9) between the Cricetulus griseus and its albino mutation strain, A:CHA, and analyze the function on the albino phenotype. Methods The total mRNA of skin tissue in the Cricetulus griseus and A:CHA was extacted. The PCR method was used to select primers. The expression levels of SLC24A5, SLC39A9 was analyzed by real time PCR. Results The ratio of expression level of SLC24A5, SLC39A9 between Cricetulus griseus and A:CHA were 2 and 3.8 times. Conclusion The gene expression level of SLC24A5, SLC39A9 in the A:CHA were lower than that in the Cricetulus griseus. The study strongly supports the notion that the two genes may be contribute to the albino phenotype.
    Refinement on Establishment of Atrial Fibrillation Model in Rabbit
    ZHANG Fang-liang, CHEN Ying-min, SONG lei, LUO Zhang-yuan, ZHANG Wen-zan, XU Xiang-dong, HE Ben
    2014, 34(6):  443-448.  DOI: 10.3969/j.issn.1674-5817.2014.06.003
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    Objective To optimize the method for establishing an animal model of atrial fibrillation(AF) based on self-designed implantable telemetry and stimulator. Method First, the anesthetic protocols were refined. Thirty New Zealand rabbits, half males and half females, were divided into three groups. Group A, pentobarbital sodium only, Group B, xylazine with diazepam and lidocaine, and Group C, xylaxine with midazolam and lidocaine. Second, the location of recording electrodes were optimized by using a needle to connect to ECG collect electrode and the needle was punctured to the different location of the rabbit chest subcutaneous to analyze the electrocardiogram before choose the best location. Third, the stimulating electrode was refined and the method of operation was improved. Results At the anesthesia, the heart rate and breath rate of Group A were depressed more than Group B and Group C, first anesthesia time of Group B was longest, complemental anesthesia times of Group C was more than Group A and Group B. The best electrode location could prove good ECG record . Refined stimulating electrode could improve the effect of stimulation. Reformed operation method could simplify the operation procedure. Conclusion The refined method for establishing an animal model of atrial fibrillation used by lmplantable telemetry and stimulator was feasible, and more conformance to animal welfare.
    Effect of Metallothionein and Rosuvastatin on Atherosclerosis and SM22α Expression in ApoE-deficient Mice
    ZHU Zhong-sheng, LU Ya-li
    2014, 34(6):  449-453.  DOI: 10.3969/j.issn.1674-5817.2014.06.004
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    Objective To research the effect of metallothionein(MT) and rosuvastatin on atherosclerosis and smooth muscle 22 alpha (SM22α) expression in ApoE-deficient mice. Methods Thirty 6-week-old ApoE- deficient male mice were divided into hyperlipidemia model group, rosuvastatin group, and MT group randomly, fed with fatty diet for 13 weeks. Ten background strain C57BL/6J (wild type) male mice were selected as normal control group,and fed with normal diet for 13 weeks. After 13 weeks, eyeballs were extracted and blood was taken from ApoE-deficient mice for detecting lipid profile, then the mice were executed and the aortas were dissociated, 5 mm vessel in aorta arch were taken and fixed in 10% formalin, then treated with paraffin imbedding, slicing, HE staining and SM22α immunohistochemistry staining; some specimen was preserved in -80 ℃ refrigerator, total protein and RNA were extracted, West Blot and RT-PCR were taken.To observe morphology changes of aortic wall, to analyse SM22α expression intensity of aorta tissue qualitatively and quantitively. Results The blood lipid level in rosuvastation group was lower than that in hyperlipidemia model group (P<0.05). There is no difference in the blood lipid level between MT group and hyperlipidemia model group (P>0.05). Compared with normal control group, intimal thickness, plaque volume increased and SM22α protein and mRNA expression decreased in hyperlipidemia model group. Compared with hyperlipidemia model group, intimal thickness, plaque volume decreased and SM22α protein and mRNA expression increased in MT and rovustatin group. Conclusion The MT has no effect on lipid profile, rovustatin can lower blood lipid obviously. MT and rovustatin can alleviate atherosclerosis lesions and increase SM22α protein and mRNA expression in aorta of ApoE-deficient mice. Their anti-athrosclerosis effect is related to up regulating SM22α expression.
    Study on Comparative Histology of Digestive Glands in 6 Species of Laboratory Animals
    XU Wen-mang, LI Xia, HE Zhan-long, LI Tao, ZHAO Xi-long, WANG Yuan-yuan, DAI Fang, LIU Qiang-gao, CAI Lin, PAN Xin-yan, YANG Ju-lun
    2014, 34(6):  454-462.  DOI: 10.3969/j.issn.1674-5817.2014.06.005
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    Objective To study the histological differences on digestive glands among 6 species of laboratory animals, and observe their spontaneous lesions, providing a theoretical basis for pathological testing standards of laboratory animals. Methods 30 rhesus monkeys, 16 Beagle dogs, 18 Japanese white rabbits, 20 SD rats and 20 KM mice for study. The same and different histological structure characters of them were summarized by comparative study, spontaneous lesions of digestive gland were observed. Results (1) The major histological differences of parotid gland, submandibular gland and sublingual gland were acinar structures, adipose tissue content in acinus, AB/PAS staining status of glutinous acinus and the histological structures of conduct pipe. (2)Beagle dog had a couple of glandula zygomaticas, and Japanese rabbit had a couple of suborbital glands as a exclusive salivary gland.(3)The major histological differences of liver were the histological structures of hepatic lobule and portal area, morphological characters of hepatocytes. (4)Whether the presence of lymphoid tissue in glandular lobule and the histological structures of conduct pipe were major histological differences of pancreas. (5)Whether the presence of mucous membrane antrums and the morphological characters of germinal epithelium were major histological differences of gallbladder.(6) Inflammation was a common spontaneous lesions of liver. Conclusion Some valuable digestive gland histological data in 6 species of laboratony animals were obtained, and their different histology characters were revealed. The histological differences on digestive glands and spontaneous lesions in different animal species should be fully considered in biomedical researches.
    Preliminary Establishment of Shared Database about Comparative Histology in 6 Species of Laboratory Animals
    CHEN Ying, PAN Hua, ZHOU Guang-xing
    2014, 34(6):  463-466.  DOI: 10.3969/j.issn.1674-5817.2014.06.006
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    Objective To establish a comparative histology database of commonly used laboratory animals. Methods All the main organs and tissues from 6 species of laboratory animals, the mouse, rat, rabbit, guinea pig, dog and cynomolgus monkey, were collected. After embedded by paraffin, sectioned and stained with hematoxylin and eosin, tissue sections were observed under microscope and described. The comparative pathology database were established by SQL Server 2008 database management system. Results Tissues were compared among these 6 species of laboratory animals and distinctive pictures were got plused with descriptions. A user-friendly, inquiry conveniently and shared comparative histology database were established. Conclusion The establishment of comparative histology database are benefit for research of animal pathology and disease model in the field of bio-medicine.
    Impact of Several Welfare Intervention Measures on Stress-related Index in Mice
    ZHOU Jia-liang, HU Ying, SHA Xiao-min, YANG Fei, AI Xiao-jie
    2014, 34(6):  467-472.  DOI: 10.3969/j.issn.1674-5817.2014.06.007
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    Objective To study the impact of frequently-used material intervention measures towards stress-related index of laboratory animals. Methods Sixty weaning male KM mice were randomly assigned to five groups: A was routine feeding group, B nutritional intervention group, C sunflower seed group, D toy group, and E incense group, with 12 mice in each group. Food and water intake were measured every day. Body weight was measured every two days. Blood sample was collected 37 days later from the caudal vein in mouse tail and blood routine examination has been performed. mRNA index of GR, POMC, nNOS, BDNF, NMDA, TH and TPH in brain tissues of the mice were measured by fluorescent real-time PCR method and differences between material intervention groups and routine feeding group were compared. Results Compared with group A, the body weights of mice in other groups reduced. For food intake, group B reduced compared with group A, while group D and E increased. For water intake, group B reduced compared wtih group A, while group D and E increased. For the level of BDNF mRNA expression, comparing with group A, group F and E increased obviously, while group C reduced to some extent. For the level of GR mRNA expression, comparing with group A, group D reduced obviously, while group C and E reduced to some extent. For the level of POMC mRNA expression, comparing with group A, group D reduced to some extent, while other groups reduced significantly. For the level of NMDA mRNA expression, comparing with group A, group D increased obviously. For the level of TH mRNA expression, comparing with group A, group B and C reduced to some extent. For the level of TPH mRNA expression, comparing with group A, group C reduced to some extent. Hematological parameters had no obvious difference among all groups. Conclusions Placing toys makes food and water intake rise sharply, ability to resist fear and anxiety enhanced obviously, learning and memory ability reinforced greatly and stress-related index reduced evidently. Incense makes food and water intake go up, ability to resist fear and anxiety enhanced obviously and stress-related index reduced evidently .
    Microphthalmia Mutagenesis Mouse Induced by N-ethyl-N-nitrosourea and Its Genetic Tests
    WANG Jian-ming, GENG Teng, CHEN Hang, NI Jun-da, WANG Wen-hua, CHEN Bing, XUE Zheng-feng
    2014, 34(6):  473-475.  DOI: 10.3969/j.issn.1674-5817.2014.06.008
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    Objective To obtain the eye mutation mice induced by ethylnitrosure (ENU) which would offer the models for human eye diseases research. Methods N-ethyl-N-nitrosourea (ENU) was intraperitoneally injected in forty C57BL/6J (B6) male mice aged 8-10 weeks old. The male mice were mated with the same strain female mice, and their progenies were screened for visible eye mutation. Genetic tests were carried out. Results An inherited microphthalmia mouse was obtained by ENU-induced mutagenesis. Genetic tests showed that genetic background can affect the penetrance of the defect. The penetrance of observed eye defect were 90%, 0% and 27.8% on the B6, BDF1 and BDBN2, respectively. Conclusion The microphthalmia mice models was obtained by ENU-induced mutagenesis, it may be used for human microphthalmia disease research.
    Analysis and Disposal of Diarrhea and Wasting in Common Marmosets
    LI Na, WANG Wen-yu, HE Wei, DING Shuang-shuang, XI Jian-fei, XU Hao, ZHANG Zhou
    2014, 34(6):  476-480.  DOI: 10.3969/j.issn.1674-5817.2014.06.009
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    Objective To find out the cause of diarrhea and wasting in common marmosets (Callithrix jacchus), and formulate feasible prevention and control actions, which can provide experience for large-scale marmosets breeding. Methods The causes of diarrhea and wasting were found, by analyzing symptoms, diet, feeding, pathologic autopsy, tissue section, anal swab inspection, etc. Results By using laboratory diagnosis and care management improvement, the diarrhea and wasting in marmosets was due to the changes in housing environment such as unstable temperature, breeding under the air-condition outlet, or frequent entrances and exits of staff which activated conditional pathogenic bacteria Klebsiella pneumoniaeand and led to two deaths. The disease was controlled gradually by changing the way of feeding management. Conclusion Scientific management and good veterinary care play the important role in prevention of the diarrhea and wasting in common marmosets.
    Selection of Modeling Drugs for Assessment of Facilitating Feces Excretion Function
    SUN Yu-li, PAN Jing-yi, WANG Li-ping, PAN Jie, YANG Jun, HONG Xin-yu
    2014, 34(6):  481-485.  DOI: 10.3969/j.issn.1674-5817.2014.06.010
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    Objective To evaluate the validity and rationality of loperamide as the modeling agent to asses the effect of relaxing bowel using diphenoxylate as a reference. Methods The appropriate dosages of diphenoxylate and loperamide as modeling agents were screened. The validation tests were performed with the same health food which had been sold as having the facilitating feces excretion function. The small intestine movement experiment and the defecation experiment were carried out. Results (1) Screening tests: Within the screening doses, the ink propulsion rate of the two modeling drugs were less than the control group (P<0.05), so both of the small intestine peristalsis inhibition were established successfully. Within the screening doses, loperamide at the dose of 9 mg/kg and 10 mg/kg induced the constipation, none of diphenoxylate. (2) Validation tests: The animal models were succeeded with the same dosage of diphenoxylate and loperamide. The ink propulsion rates of the test substance were greater than their model control groups respectively (P<0.01). The first defecating time, the amount of feces and fecal weight within 6h of the test substance were significantly different with their model control groups (P<0.01). Loperamide and diphenoxylate have the same effect of functional food. Conclusion Loperamide can be used as amodeling drug for assessing the facilitating feces excretion function. in the appropriate dose.
    Comparison on Rat Liver Tissue Slides Embedded by Paraffin among Three Different Fixatives
    ZHAO Yu-qiong, CHEN Hua
    2014, 34(6):  486-489.  DOI: 10.3969/j.issn.1674-5817.2014.06.011
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    Objective To investigate the difference of three kinds of conventional fixatives on rat liver tissue slides embedded by paraffin and stained with HE. Methods Three kinds conventional fixatives, 10% neutral buffered formalin, Bouin's solution and modified Davidson's solution, were used to fix rat liver tissues with the animals in fasting or non-fasting conditions. The fixed tissues were embedded by paraffin, sliced and stained with HE. The qualities of the tissue slice were compared among different fixatives fixed under different conditions (fasting or non-fasting). Results Hydropic degeneration was observed in non-fasting rat liver tissue, and the liver tissue structure was fuzzy. The slices fixed by Bouin's solution and modified Davidson's solution showed clearer tissue structures, but not as brightly as the slices fixed by 10% neutral buffered formalin fixatives. Conclusion The conditions of fasting or non-fasting affect the liver tissue structure greatly. However, there is no significant difference among three kinds of fixatives fixed slices.
    Non- pharmaceutical Pentobarbital Preparation and Anesthetic Effect Evaluation
    LI Cai-Yun, PAN Xue-ying, LUAN Han-Sen, ZHANG Chao, SONG Zheng, CHANG Yan
    2014, 34(6):  496-499.  DOI: 10.3969/j.issn.1674-5817.2014.06.013
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    Objective In order to make the anaesthetic-pentobarbital that has been using on laboratory animals in domestic to meet international standard, established the preparation method for non-pharmaceutical grade sodium pentabarbital; Evaluated anesthetic effect of SD rat for three different solvent formulations. Methods Three different solvent systems were formulated 30mg/ml and 50 mg/ml pentobarbital sodium injection respectively, and tested dissolubility, pH, Conc. and the stability stored at room temperature and 2 ~ 8 ℃ of each pentobarbital sodium injection; Pentobarbital preparation with three different solvent are injected in SD rats, colleted valid duration of anesthesia and anesthetic state to evaluate anesthatic effection. Result The dissolubility of pentobarbital is great when prepared by Ethanol-Propylene glycol and HS15; The pH is stable on 9.6±0.2; the solution is very stable at room temperature and 2~8 ℃ during 30 days; For anesthetic effect evaluation of pentobarbital sodium injection in SD rats, the ethanol- propylene glycol group has short anesthesia time, minor individual differences, and the most animals reaching up to the ideal anesthetic effect. Conclusion The ethanol-propylene glycol as a solvent for the solubility and stability of the non-pharmaceutical grade sodium pentobarbital are very good, meanwhile it has the best animal anesthetic effect; the HS15 group followed.
    Discussion on the Scientific Basis of Taking NOEV[dB(C)] as the Index and Limitation Value of Noise in Laboratory Animal Facility
    WEI She-lin, YUAN Wen, CAI Qiao-bing
    2014, 34(6):  507-508.  DOI: 10.3969/j.issn.1674-5817.2014.06.016
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    The index of noise is dB(A) in the national standard for laboratory animal facility and environment of P. R. China GB14925-2010 and in relative standards of many foreign countries. This index is based on the character of human being hearing. The limitation value of the noise in standard of P. R. China is ≤60 dB(A). That is also from the demand of human being hearing. But many natural evidences and scientific researches show that the index and limitation value is not reasonable nor scientific. The writers discussed such argument based on available scientific documents. And the writers proposed that NOEV[dB(C)], No observed Effect Volume [dB(C)], could be the suitable index and limitation value for noise in laboratory animal facility and environment based on available scientific documents, and derived that NOEV[dB(C)] is different between different laboratory animals. The detailed data could be obtained by new scientific researches. The revivew pointed out the direction for how to get the limitation value of noise in laboratory animal facility and environment.
    Research Progress on Sendai Virus against Tumors
    ZHANG Quan, ZHU Hui-xia, LI Ling-yu, TAN Hai-ming, XUE Zheng-feng
    2014, 34(6):  515-518.  DOI: 10.3969/j.issn.1674-5817.2014.06.019
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    Sendai virus is one of the common pathogens in laboratory rats and mice. Transplanted tumors may be inhibited by type I interferon induction, immune response regulation and directly apoptosis in cancer cells after Sendai virus infection in hosts. Nowadays, Sendai virus has already been gradually developed as an effective and new anti-cancer reagent. In this review, the mechanisms of Sendai virus infection against cancer growth and interference on experiment were described.